Mechanism of the epidermal growth factor receptor in promoting endothelial cell dysfunction in gestational diabetes mellitus

Figure 4 EIF4A3 binds to long noncoding RNAs H19 to increase the stability of epidermal growth factor receptor messenger RNA. A: The expression of EIF4A3 was detected via quantitative real-time polymerase chain reaction (RT-qPCR); B: The expression of EIF4A3 in mouse placental tissues was detected via RT-qPCR; C: RNA pull-down and Western blotting showing the significant enrichment in EIF4A3 pulled down by H19; D: RNA immunoprecipitation verified the binding of H19 to EIF4A3; E: Western blotting showing the knockdown efficiency of EIF4A3; F: RT-qPCR detection of the expression of epidermal growth factor receptor (EGFR) messenger RNA (mRNA); G: EGFR expression detected by Western blotting; H: H19 expression detected by RT-qPCR; I: H19 overexpression efficiency detected by RT-qPCR; J: 5 μg/mL actinomycin D (ActD)-treated cells for 0, 4, 8, and 12 hours, followed by overexpression of H19 to detect the expression of EGFR mRNA; K: Cells treated with 5 μg/mL ActD for 0, 4, 8, and 12 hours, and EGFR mRNA expression was measured after EIF4A3 knockdown; L: The expression of EGFR mRNA in cells was detected via RT-qPCR. ^aP < 0.05. ^bP < 0.01. ^cP < 0.001. mRNA: Messenger RNA; NC: Normal control; HG: High glucose; GDM: Gestational diabetes mellitus; Bio-H19: Biotinylated H19; Bio-NC: Biotinylated negative control; sh-NC: Short hairpin-negative control; sh-EIF4A3: Short hairpin RNA targeting EIF4A3; EGFR: Epidermal growth factor receptor; GAPDH: Glyceraldehyde-3-phosphate dehydrogenase; OE-NC: Overexpression negative control; OE-H19: Overexpression H19; sh-H19: Short hairpin RNA targeting H19.