Figure 4.

LSD1 inhibitor pargyline (PG) treatment attenuates kidney cancer growth and induces cell death additively with enzalutamide. (A) Caki-2 cells were treated with enzalutamide and/or the indicated concentrations of PG and the optical density values of viable cells were determined using EZ-Cytox solution. Bars represent the means ± SD of three independent experiments, and * denotes p < 0.05 (Student’s t-test) versus enzalutamide-only treated group. (B) After two days of treatment with the indicated drugs, the total cell lysates were collected and immunoblotted with the indicated antibodies. (C) Flow cytometry analysis of annexin V and propidium iodide (PI) staining of apoptotic cells following treatment of Caki-2 cells with the indicated drug for three days. The percentage of early apoptosis (EA) and late apoptosis (LA) cells are calculated and graphed below. Error bars represent the mean ± SD of three independent experiments and * denotes p < 0.05 (Student’s t-test) between the two indicated groups. (D) The mRNA levels of indicated genes in Caki-2 cells treated with enzalutamide and/or pargyline were analyzed by RT-qPCR. Error bars represent the mean ± SD of three independent experiments and * denotes p < 0.05 (Student’s t-test) between the two indicated groups. (E) Western blots of whole-cell lysates after 24 h of treatment with the indicated drugs and concentrations. Whole-cell lysates were immunoblotted with indicated antibodies.