Figure 2.
Senescent idiopathic pulmonary fibrosis (IPF) patient-derived lung fibroblasts express spontaneously elevated Pim-1 and p65/RelA phosphorylation. A: lung fibroblasts from non-IPF or IPF donors were cultured for 24 h in the absence of serum before total cell protein isolation and Western Blot analysis. n = 3 non-IPF and 4 IPF biologically independent samples (**P < 0.01 vs. non-IPF). B: lung fibroblasts from non-IPF or IPF donors were cultured for 24 h in the absence of serum before RNA isolation and qPCR analysis. RNA transcript levels are quantified relative to the same representative non-IPF sample, using GAPDH as a housekeeping gene. n = 4 non-IPF and 7 IPF biologically independent samples (*P < 0.05, **P < 0.01 vs. non-IPF). C: lung fibroblasts from non-IPF or IPF donors were stained for senescence-associated β-galactosidase (SA-β-gal) staining as a marker of cellular senescence. Representative examples for non-IPF and IPF are shown.