Fig. 6: Modulation of THBS1 enhances the healing capacity of late irradiated human skin. | Nature Communications

Fig. 6: Modulation of THBS1 enhances the healing capacity of late irradiated human skin.

From: Epigenetic memory of radiotherapy in dermal fibroblasts impairs wound repair capacity in cancer survivors

Fig. 6

A Illustration of THBS1 sgRNA design and qRT-PCR analysis of THBS1 expression in human fibroblasts transfected with CRISPR/Cas9-SAM plasmids (n = 3). qRT-PCR of ECM genes B–E and ACTA2 (F) in fibroblasts with THBS1 overexpression (OE). (B, C) Ctr, n = 7, OE, n = 5; (D) Ctr, n = 7, OE, n = 4; (E) Ctr, n = 6, OE, n = 5; (F) Ctr, n = 7, OE, n = 6. G Scratch wound assays of fibroblasts with THBS1 OE (n = 7). Representative images of wounds are shown: black and white dashed lines indicate the wound edges at 0 and 24 hours, respectively. Scale bars: 300 µm. qRT-PCR analysis of THBS1 (H) and ACTA2 (I) in RT+ and RT− fibroblasts with THBS1 silencing and TGF-β1 treatment (n = 3). J Scratch wound assays of RT+ fibroblasts with THBS1 silencing and TGF-β1 treatment (siCtr, n = 5, siTHBS1, n = 4). Representative images of wounds 18 hours after scratching are shown. Scale bars: 300 µm. K Scratch wound assays of fibroblasts with THBS1 OE and treated with or not with ERK inhibitor (U0126). Representative images of wounds 12 hours after scratching are shown (n = 7). Scale bars: 300 µm. L Representative images of ex vivo wounds on RT+ skin treated with or not with anti-THBS1 antibodies (Control, n = 5; THBS1 antibody, n = 4). Wound contraction was evaluated by measuring the change of areas within the initial wound edges (indicated with dashed lines) on days 0-6 post-injury. Scale bars: 500 µm. qRT-PCR of RUNX1 M), COL3A1 (O), and ELN (P) in RT− and RT+ fibroblast transfected with siRUNX1 and treated with or not with TGF-β, n = 3 M), n = 4 (O, P). N Scratch wound assays of RT− and RT+ fibroblasts transfected with siRUNX1 (RT− or RT+ with siCtr, n = 5; RT− or RT+ with siTHBS1, n = 9). Representative images of wounds 10 hours after scratching are shown. Scale bars: 300 µm. Data are presented as means ± SD. One-way ANOVA (A), unpaired two-tailed student’s t-test (B–G, J, M), two-way ANOVA (H, I, K, L, N–P). ns: not significant. Source data are provided as a Source Data file.

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