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. 2019 Nov 5;12:9143–9152. doi: 10.2147/OTT.S226796

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© 2019 Cai et al.

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Effects of METTL3 depletion on SHH-GLI pathway and apoptosis. (A)The protein level of SHH-GLI pathway was analyzed by Western blotting after METTL3 depletion. (B, C) The mRNA level of c-Myc and cycling D1 was analyzed by qRT-PCR. *P ˂ 0.05, **P ˂ 0.01. (D, E) The protein level of Bak, Bax, Bcl-2, Bcl-xL, and cleaved PARP was analyzed by Western blotting after METTL3 depletion. (F) The relative caspase-3/7 activity was measured using Apo-One™ homogenous caspase-3/7 assay. **P ˂ 0.01. (G, H) LNCaP and PC3 cells were transduced with Scr or METTL3 shRNA for 48 hrs followed by overexpression with wild type or mutant METTL3 plasmid for 24 hrs. GLI1 protein level was detected by Western blotting (G) and the methylated GLI1 mRNA level was analyzed by Me-RIP assay (H). *P ˂ 0.05.