Fig. 4

RNF113A functions as an E3 ubiquitin ligase to mediate Circ_0001187-induced METTL3 degradation. A Western blot results of METTL3 in THP-1 cells transduced with the siRNA of potential E3 ligases respectively compared with negative control. B The expression levels of 9 potential E3 ligases in THP-1 cells after downregulating Circ_0001187 with siRNA through qRT-PCR. C Expression levels of RNF113A in AML patients and healthy controls through qRT-PCR. D The results of correlation between the expression level of RNF113A and Circ_0001187. E The effect of si-Circ or si-Circ/oe-RNF113A on the apoptosis of THP-1 and Molm-13 cells cultured with Ara-C (1 μM, 24 h). F IF assays showing the co-localization of RNF113A with METTL3 in THP-1 and Molm-13 AML cells. Scale bar, 20 μm. G Co-IP showed the binding of METTL3 with RNF113A in 293 T cells. H Western blot results of METTL3 in 293 T cells with the RNF113A knockdown or overexpression treated with DMSO or MG132 (10 μM, 6 h). I The results of METTL3 ubiquitination in 293 T cells expressed GFP-RNF113A, Flag-METTL3 and HA-ubiquitin and treatment with or without MG132 (10 μM, 6 h). J Western blot results of investigate the form of polyubiquitin chains linked to METTL3. K The results of RNF113A-induced METTL3 ubiquitination detected in WT HA-UB and K48 transfected cells. L The effect of RNF113A overexpression or RNF113A/METTL3 overexpression on the proliferation of THP-1 and Molm-13 cells. M The effect of RNF113A overexpression or RNF113A/METTL3 overexpression on the apoptosis of THP-1 and Molm-13 cells cultured with Ara-C (1 μM, 24 h). *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001; ns: Not significant