Fig. 4

In vitro studies revealed that HHLA2 overexpression could reverse the inhibition of cellular functions mediated by METTL3 depletion. We further constructed the HHLA2 overexpression cellular models based on the METTL3 depletion in 786-O and ACHN cells. LV-NC, LV-METTL3-shRNA, LV-NC + LV-HHLA2^OE, and LV-METTL3-shRNA + LV-HHLA2^OE cells were involved in the present study. A and B Based on the qRT-PCR results, the expression of METTL3 at the mRNA level was significantly decreased after METTL3 depletion, and HHLA2 mRNA expression level was significantly increased after HHLA2 overexpression upon METTL3 depletion. C Based on the Western blot analysis, METTL3 protein expression was significantly decreased after METTL3 depletion, and HHLA2 protein expression level was significantly increased after HHLA2 overexpression upon METTL3 depletion. D In 786-O cells, after depletion of METTL3, overexpression of HHLA2 could significantly increase the cellular viability. E In ACHN cells, after depletion of METTL3, overexpression of HHLA2 could significantly increase the cellular viability. Un-paired t test and Two-way ANOVA analyses were used respectively. ^*P < 0.05, ^**P < 0.01, ^***P < 0.001, ^****P < 0.0001