Fig. 5

Male C57BL/6 mice subjected to bleomycin injections on alternate days show higher dermal collagen content than male C57BL/6 mice given daily bleomycin injections. a Representative images of H&E-stained sections of male C57BL/6 mouse skin treated with subcutaneous NaCl or bleomycin injections given either daily or every other day over a period of 3 weeks. Original magnification ×100. b Dermal thickness of male C57BL/6 mouse skin treated with subcutaneous NaCl or bleomycin injections given either daily or every other day over a period of 3 weeks. Four high-power field images were taken (two measurements per image). Results represent the relative fold-change compared to NaCl-treated control mice. c Representative images of Masson’s Trichrome-stained sections of mouse skin harvested after 3 weeks of treatment with NaCl or bleomycin injected either every day or every other day. Original magnification ×100. d Hydroxyproline assay which was used as an indicator of collagen content in mouse skin treated with bleomycin every day vs alternate days. Results are represented as means ± standard error of the mean (SEM) of triplicate measurements obtained from six mice (two biopsies per mouse) and shown as relative fold-change compared to NaCl-treated control samples. e Representative images of alpha smooth muscle actin (α-SMA) immunohistochemical analysis. Original magnification ×200, inset ×630. f Relative number of α-SMA-positive cells in the dermis of mice treated with NaCl or bleomycin either every day or alternate days. Results represent the relative fold-change compared to NaCl-treated control mice. All values represent means ± SEM; n = 6 in each group