Figure 1
From: Ketotifen directly modifies the fibrotic response of human skin fibroblasts
αSMA gene expression and protein levels are reduced in TGFβ1-activated fibroblasts following ketotifen treatment. (A, B) HDFa and (C, D) WS1 fibroblasts were treated for 48 h with DMEM supplemented with 10% FBS as mock, 10 μM or 25 μM ketotifen, 10 ng/mL TGFβ1, or 10 ng/mL TGFβ1 with ketotifen added during the final 24 h. Gene expression of αSMA (ACTA2) was measured using RT-qPCR and normalized to housekeeping gene HPRT. HDFa cells were treated for 48 h under the same conditions and probed for αSMA protein by western blot. (E) A representative image of the blots is shown (left) and semi-quantitative assessments plotted (right) using GAPDH as loading control. Full-length blots are available in Supplementary Fig. 4. (F) COL1A1 was measured in HDFa cells treated with conditions using 25 μM ketotifen. Protein levels of pro-collagen 1α1 (G) and fibronectin (H) were determined by ELISA staining kits. Data shown as mean ± SEM. n = 3–6 per treatment condition. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001. αSMA alpha-smooth muscle actin, GAPDH glyceraldehyde 3-phosphate dehydrogenase, HDFa human dermal fibroblasts (adult), SEM standard error of the mean, TGFβ1 transforming growth factor-beta.
