(A) Representative images showing WNT5A-induced changes in F-actin, Vimentin filaments, and microtubules with (B) quantification of each cytoskeletal component in relation to its subcellular localization. (C) Violin plots showing the force required to rupture magnetic beads coupled with LAP-TGF-β1 peptides from dermal fibroblasts incubated with WNT5A with or without the actin inhibitor cytochalasin D (n ≥ 50 for each group). (D) Levels of active TGF-β in the supernatant of dermal fibroblast incubated with WNT5A in the presence or absence of cytochalasin D measured by TMLC assays (n = 6 for each group). (E) Representative Western blots showing the levels of P-SMAD3 in dermal fibroblasts stimulated with WNT5A and cytochalasin D (n = 3 for each group). The black thin vertical lines were drawn to separate noncontiguous lanes. (F) Volcano plot and (G) heatmap illustration of DEGs from RNA-Seq of WNT5A-stimulated human dermal fibroblasts treated with Cytochalasin D or with vehicle. (H) Deenrichment of GO biological processes related to fibroblast-to-myofibroblast transition and fibrosis (n = 3 for each group). (I) Representative Trichrome stainings, and (J) quantification of the dermal thickness, hydroxyproline content, myofibroblast counts, and active TGF-β in skin lysates from mice with WNT5A-induced skin fibrosis with or without ITGAV inhibitor (n = 5 for each group). Results are shown as median ± IQR with data representing individual data points. The statistical significance was determined by 1-way ANOVA with Tukey’s multiple comparison test. Adv, Adenovirus; Actin-i, Actin inhibitor.