Differential gene expression in heterophils isolated from commercial hybrid and Thai indigenous broiler chickens under quercetin supplementation

Figures & data

Table 1. Sequence of the primers used in real-time quantitative RT-PCR.

No.	Gene name	Accession no.	Primer	Sequence	Annealing temp (°C)	Product size (bp)
1	IL1B	NM_204524	Forward	CGC TCA CAG TCC TTC GAC ATC	60	230
			Reverse	CCG CTC ATC ACA CAC GAC ATG T
2	IL6	NM_204628	Forward	TGT TCG CCT TTC AGA CCT ACC	60	123
			Reverse	TTA TCA CCA TCT GCC GGA TCG
3	IL8L1	NM_205018	Forward	CCT CAC TGC AAG AAT GTG GAA G	60	124
			Reverse	TAT TGG TGT CTG CCT TGT CCA G
4	IL8L2	NM_205498	Forward	TGG TTT CAG CTG CTC TGT CG	60	193
			Reverse	AAG CAC ACC TCT CTT CCA TCC
5	IL12B	NM_213571	Forward	GTG GCT CGC ACT GAT AAA TCT G	60	218
			Reverse	CAG GTC ACA GTT CCA TTT GTG C
6	IL17A	NM_204460	Forward	TTT CTG CAC ATG GGA AGG TG	60	144
			Reverse	CCT GGT TCA TGT TGC TGA TGC
7	IL18	NM_204608	Forward	AGG AGG TGA AAT CTG GCA GTG	60	103
			Reverse	TCT TCT ACC TGG ACG CTG AAT G
8	CCL4	NM_204720	Forward	TCT CGC TCT TCT CAT CGC ATC	60	166
			Reverse	GAA GAT GAT GGC AGG CTT TGA G
9	LITAF	NM_204267	Forward	ACT GTG TAT GTG CAG CAA CC	60	169
			Reverse	AGC AAC AAC CAG CTA TGC AC
10	MIF	NM_001305091	Forward	TAC ATA GCC GTG CAC ATC GTA C	60	238
			Reverse	ATG CAA AGG TGG AAC CGT TC
11	NOS2	NM_204961	Forward	AGT TGT GCA TTG AGC TTG GG	60	211
			Reverse	TCA AGG AGC ATG TTG GCA AC
12	TGFB1	JQ423909	Forward	TGA GTA TTG GGC CAA AGA GCT G	60	101
			Reverse	ACA CGA AGA AGA TGC TGT GG
13	GAPDH	NM_204305	Forward	AAG GGC ATC CTA GGA TAC ACA G	60	107
			Reverse	TGG TCA TTC AGT GCA ATG CC

Figure 1. Real-time RT-PCR analyses of pro-inflammatory, inflammatory, and anti-inflammatory genes of hybrid commercial chickens under quercetin supplementation or PBS. Bar graphs show relative mRNA expressions of study genes. The data are represented as mean ± SE (n = 4–7 pools), *p < .05, **p < .01, ***p < .001 by unpaired Student t test.

Figure 2. Gene expressions in heterophils of Thai indigenous chicken breed (Pradu Hang Dam) under the effects of quercetin or PBS by real-time RT-PCR analyses. Bar graphs show relative mRNA expressions of study genes. The data are represented as mean ± SE (n = 8 pools), *p < .05, **p < .01, ***p < .001, ****p < .0001 by unpaired Student t test.

Figure 3. Venn diagram summarizes similarities and differences among up-regulated or down-regulated of gene expressions in heterophils supplemented with quercetin. As depicted, hybrid chickens are very different from Pradu chickens.

Figure 4. The heat map depicts a mean expression level of each studied gene in chicken heterophils from hybrid broilers (A) and from Pradu chickens (B) with either PBS or quercetin treatment. The expression values ranged from −3 to +3. Red indicates up-regulation, green indicates down-regulation, and black indicates no change in gene expression. The expression levels are according to the colour scale of the log₂ values.

Figure 5. CytoScape and GeneMANIA interaction network of known genes with co-expression, co-localization, and pathway. Dim grey nodes represent query gene lists. Salmon red nodes represent connecting genes involved in the inflammatory response to bacterial LPS pathway and cytokine activity.