The effect of the IRE1α inhibitor 4μ8C on TGFβ‐induced skin fibrosis in C57BL/6 mice as measured by the percentage of sirius red areas in the hypodermis. Ctrl (n = 6); Ctrl + 4μ8C (n = 5); TGFβ (n = 8); TGFβ + 4μ8C (n = 8). *P = 0.046.
Representative skin tissue sections stained with sirius red for collagen (top) and αSMA (bottom). Scale bars = 50 μm.
Percentage of αSMA‐positive cells in the hypodermis of mice from the experimental setup in (A). *P = 0.024.
Representative image of the area selected for laser capture microdissection (scale bars = 100 μm).
Expression levels of αSMA mRNA in samples of hypodermis obtained by laser capture microdissection. Ctrl (n = 5), Ctrl + 4μ8C (n = 5), TGFβ (n = 5), TGFβ + 4μ8C (n = 5). *P = 0.044.
miR‐150 expression levels in the same samples as shown in (E).
Immunohistochemical staining of αSMA in primary skin fibroblasts isolated from healthy mice after treatment with TGFβ ± 4μ8C. Scale bars = 20 μm.
αSMA mRNA expression in primary mouse skin fibroblasts after treatment with TGFβ ± 4μ8C. *P = 0.049, n = 4.
Agarose gel showing levels of spliced and unspliced XBP‐1 in primary mouse skin fibroblasts treated as indicated with TGFβ and 4μ8C.
BiP mRNA expression in primary mouse skin fibroblasts after treatment with TGFβ in the absence and presence of 4μ8C. *P = 0.00007 for TGFβ versus Ctrl and P = 0.00002 for TGFβ versus TGFβ + 4μ8C. n = 4.
Western blot showing CHOP protein expression in primary mouse skin fibroblasts treated as indicated with TGFβ and 4μ8C.
miR‐150 mRNA levels in primary mouse skin fibroblasts treated as indicated with TGFβ and 4μ8C. *P = 0.00031 compared to untreated control, n = 3.
‐test. Significant differences are indicated with * and exact
‐values given. Error bars indicate s.e.m.
