Figure 4
HOTAIR KD impairs lipid storage in mature adipocytes.( a) Oil red O staining (20X) of differentiated (D9) SCR and shHOTAIR G-ASCs. LipidL droplet area analysis from PERILIPIN immunostainings of differentiated (D9) SCR and HOTAIR KD G-ASCs presented as (b) individual data scatter plot and mean (***p < 0.0001, two-way ANOVA with Dunnett's multiple comparisons test;; n = 3 experiments, with ≥ 3 fields measured per experiment) and (c) distribution frequency. (d) RT-qPCR analysis of adipogenic transcription factor gene expression at D9 (mean fold difference ± SD; *p < 0.05, **p < 0.005, ***p < 0.001, two-way ANOVA with Dunnett's multiple comparisons test; n = 4). (e) Western blot analysis of fatty acid synthase (FAS), PERILIPIN1, and FABP4 in differentiating SCR and shHOTAIR G-ASCs (representative blot from n = 3; see Fig. SS7for original blots). γ-Tubulin is shown as a loading control. (f) RT-qPCR analysis of glucose- and lipid uptake-related genes, de novo lipogenesis genes and mature adipocyte markers in differentiated (D9) SCR and shHOTAIR G-ASCs (mean fold difference ± SD; *p < 0.05, **p < 0.005, two-way ANOVA with Dunnett's multiple comparisons test; n = 4). (g) Fold induction of glycerol release in response to 100 nM isoproterenol stimulation for 2 h in differentiated (D9) SCR and shHOTAIR G-ASCs (mean fold difference ± SD; *p < 0.05, **p < 0.005, two-way ANOVA with Dunnett's multiple comparisons test; n = 3).
