FIGURE 1.

Effect of KS18 on Mcl-1 in MM cells. (A, B), U266 MM cells were treated with or without specified doses of KS18 for 24 h, or with 5 µM of KS18 at the designated time intervals (B). (C), U266 MM cells were subjected to treatment with or without 5 μM of KS18 for a duration of 24 h. Subsequent to incubation, immunocytochemical analysis was conducted as outlined in the Materials and Methods section. ImageJ software was utilized to compute the integrated density of individual samples (n = 10), and an unpaired t-test was conducted using GraphPad Prism. (D), The selection of Mcl-1 shRNA or non-targeting shRNA transfected cells. (E), Western Blot was used to evaluate the silencing effect of the shRNA constructs. The non-targeting shRNA transfected cells served as control. (F), U266 and U266 Mcl-1 knockdown cells were administered KS18 (5 µM) for 24 h, after which an apoptotic test was conducted as outlined in the Materials and Methods section utilizing the Muse^® Cell Analyzer. For section (A, B, E), subsequent to incubation, the cells were lysed and analyzed via immunoblotting as outlined in the Materials and Methods section, utilizing the specified antibodies. The GAPDH antibody functioned as a loading control. Cells treated with vehicles served as the control. For section (F), GraphPad Prism was utilized to generate graphs and statistical calculations. ****P ≤ 0.0001.