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

Figure 3.

Figure 3.

CHIR 99021 suppresses GSK-3β. (A) Confluent orbital fibroblasts obtained from GO patients (n = 3) were treated with DMSO and 10 ng/mL of IL-1β or TNF-α for 15 minutes with or without pretreatment with 10-µM CHIR 99021 for 48 hours. Western blot analyses were performed to investigate the levels of inactive GSK-3β (p-GSK-3β [S9]), active GSK-3β (p-GSK-3β [Y216]), and total GSK-3β. Although the treatment with proinflammatory cytokines led to a significant increase in the levels of inactive GSK-3β (p-GSK-3β [S9]) and active GSK-3β (p-GSK-3β [Y216]), pretreatment with CHIR 99021 significantly suppressed the increase. (B) Confluent orbital fibroblasts obtained from GO patients (n = 3) were treated with CHIR 99021 for increasing lengths of time (0–48 hours) without stimulant. Increasing durations of CHIR 99021 treatment resulted in decreased p-GSK-3β (Y216) levels. Data in the columns indicate the mean density ratio ± SD, normalized to the level of β-actin in the same sample. Representative gel images are also shown. Differences between treated and untreated cells are indicated (*P < 0.05). For full-length gel images, see Supplementary Figure S2.