Analysis of global DNA methylation and epigenetic modifiers (DNMTs and HDACs) in human foetal endothelium exposed to gestational and type 2 diabetes

Figures & data

Table 1. The primers used in RT-PCR.

Name of the primer	Sequence 5’-3’
DNMT1_F	AGGGCTACCTGGCTAAAGTC
DNMT1_R	CCTCTCCATCGGACTTGCTC
DNMT3a_F	CGAGAGCAGAGGACGAGCC
DNMT3a_R	GTGTTGAGCCCTCTGGTGAA
DNMT3b_F	TGGAGCCACGACGTAACAAA
DNMT3b_R	GCATCCGTCATCTTTCAGCCTA
HDAC1_F	ATGCAAAGAAGTCCGAGGCA
HDAC1_R	ACCCTCTGGTGATACTTTAGCA
HDAC2_F	GGGAATACTTTCCTGGCACA
HDAC2_R	ACGGATTGTGTAGCCACCTC
Beta-actin_F	AGCCTCGCCTTTGCCGA
Beta-actin_R	CTGGTGCCTGGGGCG

Table 2. List of Mean characteristics of subjects participating in the study.

Status mothers	Controls (n = 6)	GDM (n = 8)	T2D (n = 3)	P (GDM vs controls)/(T2D vs controls)
Age (years	29 ± 1.8	33 ± 5	31 ± 8	0.07/0.17
BMI	26.9 ± 1.8	33 ± 2	25.9 ± 0.6	0.06/0.2
HbA1c %	4.9 ± 0.4 (n = 3)	6.5 ± 0.3 (n = 6)	8.9 ± 0.5	0.05/0.004
RPG (mM)	4.9 ± 0.4	7.2 ± 0.7	13 ± 3	0.01/0.05
Height (cm)	160 ± 0.02	155 ± 0.02	159 ± 0.6	0.05/0.6
Weight (kg)	70 ± 5.6	80 ± 5.9	65 ± 4	0.1/0.3

#at 12 weeks pregnancy; RPG, Random plasma glucose; BMI, body mass index, HbA1c, glycosylated haemoglobin; GDM, gestational diabetes mellitus, mean ± SEM. Number of donors donated by (n).

Figure 1. Percentage of global DNA methylation in Control-HUVECs vs GDM-HUVECs and T2D-HUVECs. ELISA was performed to determine 5mC levels. **P < 0.01 vs controls. HUVEC, human umbilical vein endothelial cells; GDM, gestational diabetes mellitus; T2D, Type 2 diabetes; C, control.

Figure 2. Expression levels of the epigenetic markers (a) DNMTs and (b) HDACs in GDM-HUVECs (n = 8) and Control-HUVECs (n = 6) were measured using Qrt-PCR. Gene expression was normalized to the mRNA level of the housekeeping gene β-actin in each sample. Results are expressed as the mean ± SEM. **P < 0.5 versus controls.

Figure 3. Expression levels of the epigenetic markers (a) DNMTs and (b) HDACs in T2D-HUVECs (n = 3) and Control-HUVECs (n = 6), were measured using Qrt-PCR. Gene expression was normalized to the mRNA level of the housekeeping gene β-actin in each sample. Results are expressed as the mean ± SEM. **P < 0.01 versus controls.

Data availability statement

No large sets of sequencing or proteomic data were generated. All the data that support the findings of this study are included in the manuscript.