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. 2020 Jul 31;61(8):51. doi: 10.1167/iovs.61.8.51

Figure 6.

Figure 6.

GSK-3β mediates adipogenic differentiation of GO fibroblasts. Orbital fibroblasts from GO (n = 3) patients were cultured in adipogenic medium to induce differentiation into adipocytes. (A) Oil Red O staining showed that increasing concentrations of CHIR 99021 inhibited adipogenesis in a dose-dependent manner. (B) Measurements of the optical density of cell lysates at 490 nm showed the same results. Experiments were conducted on three cell strains, and the results are expressed as the mean optical density (%) ± SD (*P < 0.05 vs. untreated controls). (C) Western blot analyses of adipogenic transcription factors PPARγ, C/EBPα, and C/EBPβ showed decreased levels in response to increasing concentrations of CHIR. The levels of mature adipocyte markers, FABP4 and Acrp30, were significantly reduced in fibroblasts treated with higher concentrations of CHIR 99021. Increasing concentrations of CHIR led to decreased levels of inactive and active GSK-3β and increased levels of inactive (np-β-catenin) and total β-catenin. The molecular weights of the detected bands are marked to the left of the bands (kDa). Data in the columns indicate the mean density ratio ± SD, normalized to the level of β-actin in the same sample (n = 3), and representative gel images are shown (*P < 0.05 vs. control fibroblasts on day 10 of adipogenesis). For full-length gel images, see Supplementary Figure S4.