Figure 3

HOTAIR KD prevents cytoskeletal remodeling during adipogenesis.( a) Phalloidin staining of F-actin in D0 SCR and shHOTAIR G-ASCs. Nuclei are stained with Hoechst. Scale bar: 10 µm. (b) Quantification of coherence in F-actin fibers alignment in D0 SCR and shHOTAIR G-ASCs (***p < 0.0001, two-way ANOVA with Dunnett's multiple comparisons test; n = 3 experiments, with ≥ 4 fields measured per experiment, 5 measurements per field). (c) Phalloidin staining of F-actin (green) and immunostaining of PERILIPIN1 (magenta) in differentiated (D9) SCR and shHOTAIR G-ASCs. Nuclei are stained with Hoechst. Scale bar: 10 µm. (d) Relative expression of ACTA2 in differentiating scrambled control (SCR) and shHOTAIR G-ASCs, normalized to SCR D0 (mean fold difference ± SD; *p < 0.05, **p < 0.005, two-way ANOVA with Dunnett's multiple comparisons test; n ≥ 3). (e) Western blot analysis (see Fig. S7for original blots) and (f) quantification of αSMA protein levels in differentiating SCR and shHOTAIR G-ASCs (mean fold difference ± SD; *p < 0.05, two-way ANOVA with Tukey's multiple comparisons test; n = 3). γ-Tubulin is shown as a loading control.