Research Paper Volume 16, Issue 18 pp 12651—12666
Unveiling senescence-associated secretory phenotype in epidermal aging: insights from reversibly immortalized keratinocytes
Figure 1. Terminating the expression of SV40T antigen with doxycycline induction in reversibly immortalized NHEK resulted in reduced mitotic ability. (A) Immunofluorescent staining of Ki67, SV40-T, F-actin and DAPI of NHEKs and reversibly immortalized keratinocytes with PBS or Dox treatment. Scale bar = 20 μm. (B, C) Statistical analysis for (A) (****P < 0.0001). (D) EdU staining analysis of NHEKs and reversibly immortalized keratinocytes with PBS or Dox treatment. Scale bar = 20 μm (****P < 0.0001). (E) Statistical analysis for (D) (****P < 0.0001, ns indicates not significant). All data are represented as mean +/- SEM. (F) CCK-8 assay for NHEKs and reversibly immortalized keratinocytes treated with either doxycycline (Dox) or PBS control (****P < 0.0001). (G) Clone formation assay for NHEKs and reversibly immortalized keratinocytes treated with either Dox or PBS control. (H) Statistical analysis for (G) (****P < 0.0001, ns indicates not significant).