Fig. 2.

SCP2 regulated abnormal cholesterol membrane trafficking affected cell proliferation, apoptosis and cell cycle in GH3 cells. a SCP2-OE GH3 cells were incubated with 10 μM itraconazole (ITR) for 48 h. The distribution of cholesterol (blue) in the different groups (Vector, SCP2-OE and SCP2-OE + ITR) were examined by filipin staining and laser confocal microscopy. Green signal, GH staining; red signal, PI nuclear staining. Scale bar, 10 μm. b SCP2-OE GH3 cells were incubated with 10 μM itraconazole (ITR) for 48 h. Total cellular cholesterol content was measured using the Cholesterol Assay Kit. Plasma membrane and intracellular cholesterol levels were quantified using the cholesterol oxidation-based method (n = 12, ± SEM). c Vector GH3 cells were treated with Mβ-CD (20 μg/ml) or the Mβ-CD/CHO complex (20 μg/ml) for 0, 24, 48 or 72 h, and cell proliferation of the four groups (Vector, SCP2-OE, Vector + Mβ-CD and Vector + Mβ-CD/CHO) at different time points was assessed by a CCK-8 assay (n = 6, ± SEM). d Vector GH3 cells were treated with Mβ-CD (20 μg/ml) or the Mβ-CD/CHO complex (20 μg/ml) for 48 h. Cell apoptosis in the four groups (Vector, SCP2-OE, Vector + Mβ-CD and Vector + Mβ-CD/CHO) was measured by flow cytometry (n = 3, ± SEM). e The cell cycle distribution in the above four groups was analyzed using flow cytometry (n = 3, ± SEM). An unpaired t-test was used to assess statistical significance. *P < 0.05; **P < 0.01; ***P < 0.001; #, not significant.