Fig. 2

Knockdown of ABCC6 leads to a senescence-like phenotype. a – For the cell cycle analysis, the cells were synchronized at the G1 phase by serum deprivation for 24 h, restimulated with serum for 24 h, and analyzed using flow cytometry after BrDU and PI staining. The percentage of control (scr-shRNA) and ABCC6 knockdown cells (ABCC6-shRNA) in G0/G1 was recorded. The values are the means ± SD of three replicates from three independent experiments. Statistical analysis was performed using unpaired Student’s t test: **p < 0.001. b – Representative images (40 × magnification) of senescence-associated β-galactosidase staining in control and ABCC6 knockdown cells. c – Quantitative analysis of positive β-galactosidase-stained cells. Data were generated from three independent experiments performed in triplicate and are shown as means ± SD. ***p < 0.001. Unpaired data were assessed for statistical significance using Student’s t test