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. 2022 Aug 3;12:984514. doi: 10.3389/fonc.2022.984514

Figure 3.

Figure 3

BIM mediated the cell death induced by a combination of MK-2206 treatment and ATXN3 downregulation in NB cells. AS cells were transfected with ATXN3 siRNAs (#1 and #2) and BE2 cells were transfected with ATXN3 siRNA #2 (marked as ATXN3 siRNA) for 16 h followed by 48 h treatment of MK-2206. (A, B) Cell confluence was dynamically detected by IncuCyte Zoom and analyzed at the end of experiment; (C, D) Cell survival was detected by CCK8 assay; (E, F) Cell apoptosis was detected by Annexin V/PI flow cytometry. Bar, SD, **, P< 0.01, ***, P<0.001, ATXN3 siRNAs + perifosine vs. control siRNA + perifosine; (G) The images of AS and BE2 cells with ATXN3 siRNAs transfection and perifosine treatment at 0 and 48 h were recorded; (H) AS and BE2 cells were transfected with ATXN3 siRNA2 (marked as ATXN3 siRNA) for 16 h, and treated with MK-2206 for 24 h, the expression of ATXN3 and BIM was detected by Western blot; (I) BIM siRNA #1 (marked as BIM siRNA) and ATXN3 siRNA #2 (marked as ATXN3 siRNA) were transfected into AS and BE2 cells alone or combination, then the cells were treated with MK-2206 for 48 h, cell survival was detected by CCK8 assay; (J, K) Cell confluence was detected by IncuCyte Zoom. Bar, SD, **, P<0.01, ATXN3 siRNA + MK-2206 + BIM siRNA vs. ATXN3 siRNA + MK-2206.