Research Paper Volume 13, Issue 18 pp 22286—22297

Figure 4. Silencing ASXL1 sensitized ACC cells to etoposide, doxorubicin and cisplatin (EDP) regimen. (A) Relative firefly-luciferase activity of FSCN1 in NCI-H295R cells where Firefly luciferase activity was normalized to Renilla luciferase activity for all samples to yield relative luciferase activity upon ASXL1 silencing or control; (A, B) Cell count detected using CCK-8 in ACC cell lines with ASXL1 overexpression by adenovirus (Av) or control (AvCON) and/or with FSCN1 silencing by shRNA; (C) Transwell assays used to detect cell invasion with Matrigel in ACC cells with ASXL1 overexpression by adenovirus (Av) and/or with FSCN1 silencing by shRNA, captured at 100×; Flow cytometry used to detect (D) cell cycle profile and (E) apoptosis in ACC cells with ASXL1 overexpression by adenovirus (Av) and/or with FSCN1 silencing by shRNA; Dose-response curves of (F) Doxorubicin, (G) Cisplatin and (H) Etoposide in ACC cells with ASXL1 or FSCN1 silencing by shRNA or control (shCON), with IC50 (μmol) in parentheses (ns = not significant; *P < 0.05; **P < 0.01).