Fig. 5: OGD/R induces METTL3-dependent Lnc-D63785 m6A methylation in neuronal cells. | Cell Death & Disease

Fig. 5: OGD/R induces METTL3-dependent Lnc-D63785 m6A methylation in neuronal cells.

From: Oxygen glucose deprivation/re-oxygenation-induced neuronal cell death is associated with Lnc-D63785 m6A methylation and miR-422a accumulation

Fig. 5

SH-SY5Y cells (a) or the primary neurons (g) were subjected to OGD/R stimulation, and then tested by MeRIP-qPCR assay of Lnc-D63785 m6A methylation. Expression of listed proteins in the stable SH-SY5Y cells with applied METTL3 shRNA (s1/s2, two different sequences) or scramble control shRNA (“shC”) was shown (b); Cells were subjected to OGD/R stimulation and then cultured for applied time periods, and relative Lnc-D63785 m6A methylation level was tested by MeRIP-qPCR assay (c); Expression of Lnc-D63785 (d) and miR-422a (e) was tested by qPCR assays, with cell death tested by LDH release assay (f). Stable SH-SY5Y cells expressing the lentiviral METTL3 expression construct (OE-METTL3-sL1/sL2, two lines) or the empty vector (“Vec”) were established, and then expression of METTL3 protein (h), Lnc-D63785 (j), and miR-422a (k) was tested by Western blotting and qPCR assays; The relative Lnc-D63785 m6A methylation level was tested by MeRIP-qPCR assay (i); l The proposed signaling cascade of this study. Expression of listed proteins was quantified and normalized to GAPDH (b and h). Data indicate standard deviation (SD, n = 5). *p < 0.05 vs. “Mock” cells. #p < 0.05 vs. OGD/R-treated “shC” cells (c–f). # p < 0.05 vs. “Vec” cells (i–k). Each experiment was repeated three times and similar results were obtained.

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