Fig. 1: Comparative assessment of proliferative and senescent characteristics of IPF Fibroblasts.

A, B Co-localization of senescence markers (p16, p21) with RAGE (AT1), SPC (AT2), and Vimentin (fibroblast) via immunofluorescence. Representative images are shown in (A). Distribution of p16+ cells and p21+ cells across three different cell types was displayed through pie charts (B). Scale bar, 50 µm. C, D Immunofluorescence detection of the DNA damage marker (γ-H2AX) and senescence markers (p53, p16, and p21) in control and IPF lung tissues. Representative images acquired via confocal immunofluorescence and the positive expression rate are shown in (C) and (D), respectively. Scale bar, 50 µm. E, F α-SMA, Vimentin, Ki67, p16, and p21 expression in normal human lung fibroblasts (NHLF) and diseased human lung fibroblasts (DHLF) measured by immunofluorescence. EdU assay was measured to assess cell proliferative ability. Representative images and statistical analyses are shown in (E) and (F), respectively. Scale bar, 200 µm. G Immunoblotting analysis showing elevated protein levels of the senescence markers p16 and p21 in DHLF compared to NHLF. H, I Increased β-galactosidase staining in DHLFs, indicating higher senescence. Scale bar, 200 µm. (Results are presented as means ± SD, n = 5, *p < 0.05; **p < 0.01; *** p < 0.001).