Expression of HBEGF, EREG, and AREG induced by Piezo1 activation in vascular endothelial cells (VECs) depends on PKC/ERK1/2 signaling pathways. (IMAGE)
Caption
(A, B) The qRT-PCR results showed increased mRNA expression of HBEGF, EREG, and AREG induced by Yoda1 in endothelial cells and was reversed with Piezo1 knockdown in HUVEC cells. (C, D) the ELISA assay results showed that increased secretion of EREG and AREG in HCMYoda1 were reversed with Piezo1 knockdown. (E) The qRT-PCR results showed the block effect of ravoxertinib (ERK1/2 inhibitor, 5 μM), adezmapimod SB 203580 (P38 MAPK inhibitor, 10 μM), SP (c-Jun N-terminal kinase inhibitor, 25 μM), and SR11302 (activator protein-1 inhibitor, 10 μM) on the expression of HBEGF, EREG, and AREG induced by Yoda1 in HUVEC and SK-Hep1 cells. (F) WB suggests that the knockdown of Piezo1 can inhibit the activation of ERK1/2 induced by Yoda1. (G, H) The WB and qRT-PCR results for non-selective protein kinase inhibitor AM2282 (200 nM) effect on activation of ERK1/2 and the expression of HBEGF, AREG, and EREG in HUVEC cells induced by Yoda1. (I, J) WB and qRT-PCR confirmed that selective protein kinase C inhibitor GO 6983 (10 μM) could inhibit the activation of ERK1/2 and the expression of HBEGF, AREG, and EREG in endothelial cells induced by Yoda1. (K) Immunofluorescence analysis revealed that Yoda1 (5 μM for 0, 5, and 10 min) could activate PKCα. (L) The schematic diagram delineating the probable mechanism of the expression of HBEGF, EREG, and AREG induced by Piezo1 activated by Yoda1. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, ∗∗∗∗P < 0.0001; two-tailed Student's t-tests. PVEC, mouse primary vascular endothelial cell; HCMYoda1, conditioned medium from HUVEC with Yoda1-activated Piezo1.
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Genes & Diseases
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