Fig. 2

Increased expression of pHSP27 by IR. A Amounts of three phosphorylated HSP27 and total HSP27 proteins in the L132 cell line after 10 Gy IR at the indicated times by representative western blots and quantifications. ^*P < 0.05, ^**P < 0.01, ***P < 0.001 vs. Control B Immunoprecipitation and immunoblotting analyses using IkBα. L132 cells transfected with either AAA or DDD at 4 h after 10 Gy IR, displayed by representative western blots and quantifications. Ratio of each protein to β-actin in all western blot data (n ≥ 3). Biologically independent samples and results are representative of independent experiments. Data are expressed as mean ± SD. C Representative immunofluorescence image of p65 translocation in L132 cells transfected with either AAA or DDD at 4 h after 10 Gy IR (magnification, 400 ×). The graph shows the quantification of p65 and DAPI co-localization. D Representative immunofluorescence image of phalloidin (green) and DAPI (blue) in L132 cells transfected with either AAA or DDD (red). Magnification, 400 × (E) The mRNA levels of il-1β, il-6, and twist in L132 cells transfected with either AAA or DDD using qRT-PCR (n ≥ 3, mean ± SD). ^*P < 0.05 vs. AAA-Control; ^#P < 0.05 vs. DDD-Control; ^†P < 0.05 and ^†††P < 0.001 vs. AAA-IR. F Sirius red- and pHSP25 (Ser86)-stained lung Sections 6 weeks after focal 75 Gy irradiation (IR) in C57BL/6 N control and HSP25 TG mice. Graphs show scores quantifying collagen deposition and pHSP25-positive cells (n ≥ 3, mean ± SD). Magnification, 12.5 and 200 × . Scale bar, 100 μm. Subsequent statistical analysis was performed with unpaired one-way ANOVA with Newman-Keuls test for multiple comparisons