Fig. 3: TRMP regulates cell proliferation and G1/S cell cycle progression.

a A549 cells were infected with lentiviruses expressing control shRNA, TRMP shRNA#1, or TRMP shRNA#2. Forty-eight hours after infection, cells were plated (day 0), and cell numbers were counted at the indicated time points. Data shown are mean ± SD of three independent experiments. *** indicates p < 0.001. The knockdown efficiency of TRMP is shown in Supplementary Figure S2A. b A549 cells were infected with lentiviruses expressing control shRNA, TRMP shRNA#1, or TRMP shRNA#2. Forty-eight hours after infection, 200 cells were plated and cultured for an additional 10 days. The colonies were then stained with crystal violet. The shown images are representative of three independent experiments. Data shown are mean ± SD (n = 3). ** indicates p < 0.01. c A549 cells were infected with lentiviruses expressing control shRNA, TRMP shRNA#1, or TRMP shRNA#2. Forty-eight hours later, cells were subjected to EdU staining. The nuclei were also visualized by Hoechst 33342 staining. The shown images are representative of three independent experiments. The percentage of proliferating cells was expressed as the ratio of EdU-positive cells to total Hoechst 33342-positive cells. Data shown are mean ± SD (n = 3). ** and *** indicate p < 0.01 and p < 0.001, respectively. d A549 cells were infected with lentiviruses expressing either control RNA or TRMP. Forty-eight hours after infection, cells were plated (day 0), and cell numbers were counted at the indicated time points. Data shown are mean ± SD (n = 3). * indicates p < 0.05. The successful overexpression of TRMP is shown in Supplementary Figure S2F. e A549 cells were infected with lentiviruses expressing control shRNA, TRMP shRNA#1, or TRMP shRNA#2. Seventy-two hours after infection, cells were subjected to flow cytometry analysis to evaluate cell cycle distribution. Data shown are mean ± SD of three independent experiments. * and ** indicate p < 0.05 and p < 0.01, respectively