Through a glass, darkly? HepaRG and HepG2 cells as models of human phase I drug metabolism

Figures & data

Table 1. Proteomic analysis of cytochrome P450 expression in HepaRG cultures, cryopreserved PHHs and human liver biopsies.

	Protein expression (fmol/µg soluble protein)
Isoform	HepaRG	PHHs	Human liver
CYP1A1	0.01 ± 0.00	0.01–0.25	0.01–0.17
CYP1A2	0.15 ± 0.03	2.0–7.7	2.4–9.0
CYP2B6	0.10 ± 0.02	1.4-22.7	0.3–1.2
CYP2C8	0.57 ± 0.03	10.0–23.2	5.0-8.2
CYP2C9	3.11 ± 0.16	32.0–38.7	9.7–19.1
CYP2C19	1.31 ± 0.07	2.4–6.6	<2.6
CYP2D6	BLQ	5.0–8.6	0.06–7.9
CYP2E1	0.44 ± 0.02	5.1–14.7	2.6–4.6
CYP3A4	2.68 ± 0.14	29.2–93.7	1.8–8.3
CYP3A5	0.11 ± 0.00	0.31–0.63	0.2–3.1

ND: not determined; BLQ: below the limit of quantitation.

Summarized from Supplementary Tables S8–S10 of Hammer et al. (2021). Fresh frozen PHHs from three donors were thawed and prepared for analysis without cultivation or treatment. Five human liver biopsies (analyzed and published previously) were included for comparison. Undifferentiated HepaRGs were obtained from Biopredic International and grown for two weeks in the absence of DMSO then differentiated using 1.0% DMSO (days 1–3) followed by 1.7% DMSO (days 4–14).

Table 2. Basal P450-dependent activities in HepG2 and HepaRG cultures compared with PHHs.

	Substrate	HepG2	HepaRG	PHH (2D culture)	PHH (suspension)	Study
Activities expressed in pmol/min/mg protein
CYP1A2	Phenacetin	ND	∼6, ∼2, ∼2, ∼3	∼7, ∼13, ∼15	ND	1
		ND	∼0.3 to ∼0.9; ∽0.06 to ∼0.4	Mean ∼26 (n = 5)	Mean ∼18 (n = 5)	2
		∼0.9	ND	∼20 at 4 h	ND	3
				∼1 at 48 h
		0.02 ± 0.03	ND	3.9 ± 2.8	ND	4
		ND	∼0.3	∼1.8	ND	5
		BLD^1	7.65, 8.33, 10.2	2.06, 1.94, 4.34	ND	6
		∼0.1	∼2 to ∼4	∼7, ∼7, ∼10	ND	7,8
	Tacrine	1.6	1.8	ND	1.5 (pooled, single batch)	9
CYP2A6	Coumarin	ND	<∼0.2; <∼0.5	Mean ∼24 (n = 5)	Mean ∼49 (n = 5)	2
		BLD	ND	16.3 ± 9.2	ND	4
CYP2B6	Bupropion	ND	∼5, ∼2, ∼1, ∼1	∼3, ∼28, 278	ND	1
		ND	∼3 to ∼8; <∼0.3	Mean ∼9 (n = 5)	Mean ∼11 (n = 5)	2
		ND	ND	∼1 at 4 h	ND	3
				∼0.1 at 48 h
		BLD	ND	2.2 ± 3.4	ND	4
		<0.04	0.3	ND	0.7 (pooled, single batch)	9
		ND	∼2	Mean ∼3.6	ND	5
				Days 1–28
		BLD	10.6, 34.4, 50.7	BLD, 1.66, 2.61	ND	6
		∼0.6	∼3 to ∼7	∼1, ∼3, ∼6	ND	7,8
CYP2C8	Amodiaquine	ND	∼0.9	∼2.4	ND	5
CYP2C9	Diclofenac	ND	∼18, ∼9, ∼13, ∼9	∼42, ∼32, ∼72	ND	1
		ND	∼3 to ∼12; ∼0.1 to ∼5	Mean ∼26 (n = 5)	Mean ∼66 (n = 5)	2
		∼0.2	ND	∼30 at 4 h	ND	3
				∼ 0.3 at 48 h
		0.02 ± 0.02	ND	29.3 ± 14.3	ND	4
		<0.04	2.3	ND	3.1 (pooled, single batch)	9
		ND	∼5	∼6	ND	5
		BLD	3.79, 19.4, 12.5	1.81, 1.84, 3.72	ND	6
		BLD	∼30 to ∼70	∼7, ∼10, ∼8	ND	7,8
CYP2C19	Mephenytoin	ND	∼4 to ∼11; ∼0.4 to ∼4	Mean ∼10 (n = 5)	Mean ∼6 (n = 5)	2
		BLD	ND	2.5 ± 2.1	ND	4
		ND	∼0.4	∼0.8	ND	5
		BLD	2.73, 6.21, 10.3	BLD, 2.49, 0.788	ND	6
CYP2D6	Dextro-methorphan	0.02	ND	5.5 ± 7.9	ND	4
		<0.04	0.2	ND	9.6 ± 0.1 (pooled, 2 batches)	9
			ND	∼0.2	∼0.5	ND	5
	∼0.5	∼0.3 to ∼ 3	∼22, ∼11, ∼2	ND	7,8
	Bufuralol	ND	∼0.2 to ∼1; ∼0.02 to ∼0.3	Mean ∼2 (n = 5)	Mean ∼3 (n = 5)	2
		0.015	2.33, 3.34, 2.21	0.178, 0.908, 0.660	ND	6
CYP2E1	Chlorzoxazone	ND	∼0.06 to ∼0.3	Mean ∼3 (n = 5)	Mean ∼13 (n = 5)	2
		BLD	ND	12.5 ± 7.1	ND	4
CYP3A4	Midazolam	ND	∼25, ∼10, ∼2, ∼10	∼1, ∼37, ∼35	ND	1
		ND	∼3 to ∼18; ∼0.3 to ∼1.5	Mean ∼13 (n = 5)	Mean ∼ 13 (n = 5)	2
		∼0.02	ND	∼10 at 4 h	ND	3
				∼0.1 at 48 h
		<0.15	12.4 ± 2.8	ND	32.5 ± 0.7 (pooled, 4 batches)	9
		0.10 ± 0.07	ND	10.1 ± 5.2	ND	4
		ND	∼3.1	∼2.3	ND	5
		0.225	131, 114, 69.6	0.349, 33.7, 72.6	ND	6
		∼0.5	∼60	∼45, ∼43, ∼10	ND	7,8
Activities expressed in pmol/min/10^6 cells
CYP1A2	Phenacetin	ND	2.78 ± 0.728	7.18 ± 7.78	Median = 48 (n = 212)^§	10
		ND	5.00	2.82, 2.85, 9.18	ND	11
				14.9, 3.03, 19.58
		ND	4.27 ± 0.459	0.072–40 (n = 52)	1–349; median = 48 (n = 212)^§	12
				Median = 4.94
	Caffeine	BLD	0.153 ± 0.007	0.030 ± 0.003	ND	13
CYP2A6	Coumarin	ND	0.851 ± 0.159	1.03 ± 0.974	ND	10
CYP2B6	Bupropion	ND	17.9 ± 2.21	2.94 ± 2.78	Median = 32.1 (n = 137)^§	10
		ND	1.93 ± 0.0754	0.210–13.1 (n = 52)	1.86–304; median = 32.1 (n = 137)^§	12
				Median = 2.16
	Efavirenz	BLD	0.110 ± 0.007	0.18 ± 0.023	ND	13
CYP2C8	Paclitaxel	ND	0.223 ± 0.035	0.517 ± 0.802	Median = 5.24 (n = 149)	10
CYP2C9	Diclofenac	ND	3.94 ± 0.380	5.37 ± 9.54	Median = 88.2 (n = 187)	10
		ND	∼9 ± 0.7 (Day 0)	∼10 ± 7 (Day 0)	∼66 ± 14	14
			∼6 (Day 7)	∼1.5 ± 1.0 (Day 7)
			∼5 (Day 14)	∼0.5 (Day 14)
	Losartan	BLD	0.037 ± 0.003	0.005 ± 0.001	ND	13
CYP2C19	Mephenytoin	ND	1.52 ± 0.408	2.26 ± 4.71	Median = 13.8 (n = 208)	10
		ND	∼1.1 (Day 0)	∼2.2 ± 2.2 (Day 0)	∼11.3 ± 1.3	14
			∼0.7 (Day 7)	∼0.4 ± 0.7 (Day 7)
			∼1.8 ± 0.4 (Day 14)	∼0.7 ± 0.7 (Day 14)
	Omeprazole	0.053 ± 0.003	6.17 ± 0.33	0.157 ± 0.013	ND	13
CYP2D6	Dextromethorphan	ND	0.401 ± 0.062	1.18 ± 1.70	Median = 21.1 (n = 24)	10
	Bufuralol	ND	∼2.8 ± 0.3 (Day 0)	∼2.0 ± 1.6 (Day 0)	∼20.5 ± 1.8	14
			∼3.0 ± 0.1 (Day 7)	∼1.3 ± 1.4 (Day 7)
			∼3.2 ± 0.2 (Day 14)	∼0.7 ± 0.9 (Day 14)
	Metoprolol	BLD	0.230 ± 0.020	0.083 ± 0.003	ND	13
CYP3A4	Testosterone	ND	248 ± 50.9	55.2 ± 49.4	Median = 407 (n = 195)	10
		ND	∼170 ± 14 (Day 0)	∼28 ± 30 (Day 0)	∼114 ± 10	14
			∼140 ± 7 (Day 7)	∼6 ± 5 (Day 7)
			∼140 ± 4 (Day 14)	∼6 ± 5 (Day 14)
	Midazolam	ND	28.4 ± 2.39	ND	Median = 63.7 (n = 132)^§	10
		ND	38.3	7.13, 8.93, 12.73	ND	11
				16.7, 2.78, 10.67
		ND	6.67 ± 0.392	5.5–20.1	1.54–593 (n = 132); median = 63.7^§	12
0.213 ± 0.013		28.5 ± 1.50	9.13 ± 0.50	ND	13

PHH (2D culture) includes PHHs cultured on collagen-coated substrates with or without a Matrigel™ or other overlay. Where specified, PPHs were cultured for 4 – 120 h prior to assay. Approximate values (∼) were presented graphically in the original publications consulted; the values tabulated here have been estimated by eye from printed pdf versions.

^§Identical values appear to be from the same historical study. ND: Not determined. BLD: Values reported as zero are tabulated as BLD.

Studies 1-14, itemised in the right-hand column of the table, are as follows:

¹Kanebratt and Andersson (2008a). For HepaRG, the first value is in the presence of 2% DMSO, the other three are in its absence.

²Anthérieu et al. (2010). The text refers to pmol/h/mg protein, but the values are graphed as pmol/min/mg protein and these values have been used here. For HepaRG, the first range of values is in the presence of 2% DMSO, the second is in its absence.

³Ulvestad et al. (2013). Note that the results are graphed on a logarithmic scale, so it is difficult to derive accurate values.

⁴Tolosa et al. (2016). For human hepatocytes, n = 4 - 6.

⁵Kvist et al. (2018). PHHs were a single ten-donor pool.

⁶Yokoyama et al. (2018). Presents results from one batch of HepG2s, three batches of HepaRGs and three individual cultures of hepatocytes. Mean ± SD is presented in the paper, but these appear to be technical replicates rather than independent experiments.

⁷Seo et al. (2019). HepG2 values (n = 1) were assayed on Day 1; HepaRG values (n = 8) on Days 1-28. Differentiation period was from Days 15-28.

⁸Seo et al. (2020). HepaRG values (n = 1) were assayed on Day 3.

⁹Kratochwil et al. (2017). HepG2s (single value) assayed on Day 4; HepaRGs (single value) on Day 7.

¹⁰Jackson et al. (2016). CryoHepaRGs were assayed on Day 10 after thawing.

¹¹Vermet et al. (2016), values converted from nmol/h/10⁶ cells.

¹²Ramaiahgari et al. (2017).

¹³Berger et al. (2016), values converted from pmol/h/50,000 cells.

¹⁴Lübberstedt et al. (2011).

Table 3. Summary of basal P450-dependent activities in HepG2 and HepaRG cultures.

	HepG2	HepaRG
CYP1A2	Phenacetin O-deethylation activities and paraxanthine production from caffeine tend to be below or close to the limit of detection.^3,4,6,7,13 One study found phenacetin O-deethylation activity resembling that of PHHs 48 h after plating, although PHH activity was much higher 4 h after plating.^3	Phenacetin O-deethylation activities usually resemble those of PHHs in monolayer culture,^1,5,6,8,10–12 but considerably lower than historical values for PHHs in suspension.^10,12 Tacrine hydroxylation is reportedly similar in HepG2s, HepaRGs and a single batch of PHHs.^9 Paraxanthine production from caffeine may be slightly higher in HepaRGs than in monolayer cultures of PHHs.^13
CYP2A6	Coumarin hydroxylase activity is reportedly BLD.^4	Coumarin hydroxylation activity reportedly resembles that in monolayer cultured PHHs.^10
CYP2B6	Only one study reports measurable bupropion hydroxylation activity;^7 three others report activities BLD.^4,6,9 Efavirenz 8′-hydroxylation is also reportedly BLD.^13	Bupropion hydroxylation activities tend to be similar to/slightly higher than those in monolayer cultures of PHHs,^1,5,6,8–10,12 but well below the medians reported for fresh PHH suspensions.^10,12 Efavirenz 8′-hydroxylation activities reportedly resemble those of monolayer cultured PHHs.^13
CYP2C8	No reports found.	Activities toward amodiaquine and paclitaxel are reportedly slightly lower than those in monolayer cultured PHHs^5,10 and much lower than in PHH suspensions.^10
CYP2C9	Diclofenac 4′-hydroxylation and oxidation of losartan are usually close to or below the limit of detection.^3,4,6,7,9,13	Even though HepaRGs are of CYP2C9 poor metabolizer origin, diclofenac 4′-hydroxylation activities resembling those of monolayer PHHs, but considerably lower than those in PHH suspensions, have been reported.^1,5,6,8–10,14 Comparability between HepaRGs and monolayer cultured PHHs may result from loss of activity in the PHHs rather than the presence of active CYP2C9 in HepaRGs. Losartan oxidation activity, though low in both cases, is reportedly slightly higher in HepaRGs than in monolayer cultured PHHs.^13
CYP2C19	S-mephenytoin hydroxylation is reportedly BLD.^4,6 Omeprazole 5-hydroxylation has been reported at a level about 1/3 of that in monolayer cultured PHHs.^13	Similar S-mephenytoin hydroxylation activities have been reported in HepaRGs and monolayer cultured PHHs.^5,6,10,14 When compared with suspension PHHs the activities reported were considerably lower in both HepaRGs and monolayer cultured PHHs than in PHH suspensions.^10,14 Omeprazole 5-hydroxylation activity has been reported at a level about 40x that in monolayer cultured PHHs.^13
CYP2D6	Low dextromethorphan O-demethylation in has been reported.^4,9 Metoprolol hydroxylation activity is reportedly BLD.^13 Values reported for bufuralol 1′-hydroxylation in HepG2s differ by >30-fold.^6,7	Measurable dextromethorphan O-demethylation, metoprolol hydroxylation and bufuralol 1′-hydroxylation activities have been reported in numerous studies.^5,6,8–10,13,14 These often resemble those in monolayer cultured PHHs, though when compared to the activity of PHH suspensions the HepaRG activities are much lower.^9,10,14
CYP2E1	Chlorzoxazone 6-hydroxylation is reportedly BLD.^4	No reports found. Jackson et al. (2016)^10 could not quantify CYP2E1 activity in HepaRGs because of the inhibitory effect of DMSO on this isoform.
CYP3A4	Midazolam 1′-hydroxylation varies between very low values^3,4,9 and activities which are readily measurable but still ∼100-fold lower than in monolayer cultured PHHs.^6,7,13	Relatively high CYP3A4 activities are reported, whether measured as midazolam 1′-hydroxylation or testosterone 6β-hydroxylation.^1,5,6,8–14 These are usually at least as high as in monolayer cultured PHHs and often considerably higher;^6,10,11,13,14 indeed, this is the only case in which a P450-dependent activity observed in a cell line is in a similar range to that in PHH suspensions,^9,10,12,14 although the HepaRG activities still tend to be toward the lower end of the range when large numbers of PHH preparations are included.^10,12

^1–14As in Table 2.

Table 4. Studies comparing induction of CYP2B6 and CYP3A4-dependent activities in PHHs and HepaRG cultures.

CYP3A4
Inducing agent	Induction protocol and substrate	PHH	HepaRG
Phenobarbital	200 µM, 48 h, midazolam^15	ND	6.2 ± 2.4-fold
	Various concentrations, 72 h, midazolam^16	ND	Minimal at 100 µM
			∼2 to ∼15-fold at 300 µM
			∼5 to ∼51-fold at 1000 µM
	500 µM, 72 h, midazolam^17	2.97, 2.12 and 6.08-fold	36.6.-fold
		Three donors (2 M, 1 F)
	1.5 mM, 24–48 h, testosterone^18	∼1.2-fold and ∼1.3-fold (24 h)	9-fold (24 h)
			10-fold (48 h)
Carbamazepine	Various concentrations, 72 h, midazolam^16	ND	∼2 to ∼3-fold at 30 µM
			∼1 to ∼7.5-fold at 100 µM
			∼1 to ∼22-fold at 300 µM
Rifampicin	4 µM, 48 h, midazolam^15	ND	7.9 ± 2.9-fold
	Various concentrations, 72 h, midazolam^16	ND	Minimal at 0.1 µM
			∼2 to ∼10-fold at 0.3 µM
			∼7 to ∼24-fold at 1 µM
			∼9 to ∼42-fold at 3 µM
	10 µM, 72 h, midazolam^2	ND	∼6 to ∼21-fold
			on various days of culture
	(7 values with each of two substrate cocktails)
	25 µM, 72 h, midazolam^17	3.72, 2.08 and 10.6-fold	42.4-fold
		Three donors (2 M, 1 F)
	20 µM, 48 h, midazolam^19	ND	3.9-fold (2 D cultures)
			9.0-fold (tethered spheroids)
	20 µM, 72 h, midazolam^13	4.7-fold (2 D cultures)	7.9-fold
		7.2-fold (3 D cultures)
	50 µM, 48 h, testosterone^20	ND	3.8-fold (day 30); 2-fold (day 44); 2.2-fold (day 58)
	20 µM, 72 h, testosterone^14	5.2 ± 3.3-fold	2.8 ± 0.5-fold
	Various concentrations, 72 h, testosterone^10	ND	∼2.5-fold at 0.1 µM
			∼8.3-fold at 1 µM
∼14-fold at 10 µM
Hyperforin	Various concentrations, 72 h, testosterone^10	ND	∼ 2-fold at 10 nM
			∼7-fold at 100 nM
			∼7-fold at 1 µM
CYP2B6
	Inducing agent	PHH	HepaRG
CITCO	Various concentrations, 72 h, bupropion^10	ND	∼1.6-fold at 2.7 µM
			∼2.7-fold at 25 µM
			∼3.3-fold at 222 µM.
Phenobarbital	200 µM, 48 h, bupropion^15	ND	6.2 ± 4.3-fold
	1mM, 72 h, bupropion^2	ND	∼8 to ∼45-fold
			on various days of culture
	(7 values with each of two substrate cocktails)
	1.5 mM, 24–48 h, bupropion^18	∼4-fold and ∼3.2-fold (24 h)	∼ 13-fold (24 h)
			∼ 13-fold (48 h)
	500 µM, 72 h, bupropion^17	12.8 and 8.29-fold	8.66-fold
		Two donors (1 M, 1 F)	2/3 vehicle controls were BLQ
	1 mM, 48 h, bupropion^19	ND	1.9-fold (2 D cultures)
			3.2-fold (tethered spheroids)
	Various concentrations, 72 h, bupropion^10	ND	∼0.7-fold at 4 µM
			∼1.2-fold at 37 µM
∼3.7-fold at 333 µM.
Phenytoin	40 µM, 48 h, bupropion^15	ND	3.4 ± 2.1-fold
Rifampicin	4 µM, 48 h, bupropion^15	ND	5.9 ± 2.4-fold
	25 µM, 72 h, bupropion^17	8.27 and 2.50-fold	2.7-fold
		Two donors (1 M, 1 F)
	20 µM, 72 h, efavirenz^13	3.1-fold (2 D cultures)	4.0-fold
		2.7-fold (3 D cultures)

Studies on induction in PHHs are only included if they include comparisons with HepaRG, HepG2 or both. Inclusion criteria were the same as for Table 2 except that two studies which deviated slightly were included. In one of these¹⁷ microsomal preparations were used for the activity assays instead of intact cells and in the other²⁰ the detection method used was HPLC/UV rather than LC/MS. These deviations, which affect both the numerator and denominator of the induction ratio, were deemed to be unlikely to have affected fold induction values. Approximate values (∼) were presented graphically in the original publications consulted; the values tabulated here have been estimated by eye from printed pdf versions.

^1–14As in Table 2.

¹⁵Kanebratt and Andersson (2008b).

¹⁶Kaneko et al. (2009).

¹⁷Gerets et al. (2012); activities measured in microsomal preparations.

¹⁸Turpeinen et al. (2009).

¹⁹Wang et al. (2015).

²⁰Jossé et al. (2008).

Table 5. Rifampicin-induced changes in P450-dependent activities in PHHs and HepaRG cultures.

Isoform	Substrate	PHH	HepaRG
CYP1A2	Caffeine	No induction after 72 h at 20 µM in 2 D cultures; marginal induction in 3 D cultures (2.1-fold).^13	No induction after 72 h at 20 µM (1.1-fold).^13
CYP2B6	Bupropion	Using microsomal preparations, 8.27 and 2.50-fold induction of bupropion hydroxylation in PHHs from two donors (1 M, 1 F) after 72 h at 500 µM.^17	Some induction of bupropion hydroxylation is reported: 5.9 ± 2.4-fold after 48 h at 4 µM and 2.7-fold after 72 h at 25 µM.^15,17
	Efavirenz	3.2-fold induction after 72 h at 20 µM in monolayer cultures; 2.7-fold in 3 D cultures.^13	4.0-fold induction after 72 h at 20 µM.^13
CYP2C9	Losartan	Marginal, if any, induction (1.4-fold) after 72 h at 20 µM in 2 D cultures; slightly greater induction in 3 D cultures (2.6-fold).^13	No induction after 72 h at 20 µM (1.1-fold).^13
	Diclofenac	Marginal induction (1.6 ± 0.7-fold) after 72 h at 20 µM.^14	3.4 ± 0.4-fold induction after 72 h at 20 µM.^14
CYP2C19	Omeprazole	4.2-fold induction after 72 h at 20 µM in 2 D cultures; induction doubled in 3 D cultures (8.3-fold).^13	4.1-fold induction after 72 h at 20 µM.^13
	Mephenytoin	Marginal, if any, induction (1.3 ± 0.1-fold) after 72 h at 20 µM.^14	Marginal, if any, induction (1.2 ± 0.2-fold) after 72 h at 20 µM.^14
CYP2D6	Metoprolol	Marginal, if any, induction (1.4-fold) after 72 h at 20 µM in both 2 D cultures and 3 D cultures.^13	5.7-fold induction after 72 h at 20 µM.^13
CYP3A4	Midazolam	Using microsomal preparations, 3.72-fold, 2.08-fold and 10.6-fold induction is reported in PHHs from three donors (2 M, 1 F) treated for 72 h at 25 µM.^17	Most studies report moderate induction: 7.9 ± 2.9-fold after 48 h at 4 µM;^15 ∼6 to ∼21-fold after 72 h at 10 µm;^2 3.9-fold in 2 D cultures and 9.0-fold in tethered spheroids after 48 h at 20 µM;^19 7.9-fold after 72 h at 20 µM.^13 Using microsomal preparations, 42.4-fold after 72 h at 25 µM).^17 A dose-response experiment indicates induction up to ∼24-fold at 1 µm and up to ∼42-fold at 3 µM, though there was considerable batch-to-batch variability.^16
		4.7-fold induction in 2 D cultures and 7.2-fold is reported in 3 D cultures after 72 h at 20 µM.^13
	Testosterone	5.2 ± 3.3-fold induction after 72 h at 20 µM.^14	2.0, 2.2 and 3.8-fold induction after 48 h at 50 µM in cells following various periods in culture.^20
			2.8 ± 0.5-fold induction after 72 h at 20 µM.^19
			A dose-response experiment indicates ∼2.5-fold induction after 72 h at 0.1 µM, ∼8.3-fold at 1 µM and ∼25-fold at 10 µM.^10

^1–19As in Tables 2–4; this table summarizes a subset of the studies presented in Table 4.

Table 6. Cytochrome P450 mRNA expression and induction in Nuclear Receptor Knockout HepaRGs.

		PXR^− CAR^+	PXR^− CAR^−	PXR^+ CAR^−
CYP3A4	Basal mRNA expression	Increased	Decreased	Increased
	Rifampicin induction	No induction	No induction	As control
CYP2B6	Basal mRNA expression	Increased	As control	As control
	Rifampicin induction	No induction	No induction	Increased

Summarized from Preiss et al. (2021).

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