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. 2020 Nov 13;11(11):977. doi: 10.1038/s41419-020-03186-2

Fig. 4. Lentivirus-mediated overexpression or knockdown of YTHDC1 regulates neuronal survival after OGD.

Fig. 4

A Representative immunoblot of YTHDC1 in primary cortical neurons after transfection with shNC or shYTHDC1 lentivirus for 2 days. β-actin served as a loading control. B, C Cell viability (B) or LDH release (C) was assessed and quantified for transfected neurons subjected to OGD/R for 24 h or under no treatment conditions (Ctrl). The primary cultured neurons were transfected with shNC or shYTHDC1 lentivirus for 2 days before cell viability or LDH release assay. D Representative immunoblot of YTHDC1 in primary cortical neurons after transfection with LV-Vec or LV-YTHDC1 lentivirus for 2 days. β-actin served as a loading control. E, F Cell viability (E) or LDH release (F) was assessed and quantified for transfected neurons subjected to OGD/R for 24 h or under no treatment conditions (Ctrl). The primary cultured neurons were transfected with LV-Vec or LV-YTHDC1 lentivirus for 2 days before cell viability or LDH release assay. G Representative immunoblot and quantification of Bcl2 and cleaved caspase 3 (cleaved cas3) in lysates of shNC or shYTHDC1-transfected neurons subjected to OGD/R for 3 h or under normal conditions (Ctrl). β-actin served as a loading control. H Representative immunoblot and quantification of Bcl2 and cleaved caspase 3 (cleaved cas3) in lysates of LV-Vec or LV-YTHDC1-transfected neurons subjected to OGD/R for 3 h or under normal conditions (Ctrl). β-actin served as a loading control. The data are representative of three independent experiments. The data are means ± S.E.M., for all panels: *P < 0.05, **P < 0.01, ***P < 0.001 by two-way ANOVA analysis. All data are representative of or combined from at least three independent experiments.