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. 2018 Nov 19;33(3):3636–3646. doi: 10.1096/fj.201801496R

Figure 2.

Figure 2

Role of SK2 in β-cell lipotoxicity. A) INS-1 β-cells were transfected with 2 synthesized siRNAs targeting SK2 (SK2-siR1,2) and the controls (C-siR) followed by treatment with or without PA (400 μM) for 24 h. Levels of SK2 and full-length/cleaved (Clv) caspase-3 were determined by Western blotting. Numbers below lanes indicate band intensity relative to that of the loading control β-actin. The blots represent 3 independent experiments. B) INS-1 β-cells were transfected with the control siRNA (Ctl-siR), SK2-siR1, WT-SK2, or a control empty vector (EV) followed by treatment with or without PA (400 μM) for 24 h. Cell death was analyzed by flow cytometry as described in Fig. 1. C, D) SK1 (C) and SK2 (D) activity was determined, respectively, in INS-1 β-cells treated for 24 h with 400 μM PA or with vehicle alone (Veh) after pretreatment with or without 10 μM ABC294640. E, F) Cell viability was determined in INS-1 β-cells treated for 24 h with PA at increasing concentrations as indicated after pretreatment with 10 μM ABC294640 (E) or treated with 400 μM PA after pretreatment with ABC294640 at 0, 5, 10, and 20 μM (F). Data are expressed as mean ± sd (n = 3). *P < 0.001.