Beta-Arrestin 1 Mediates Liver Thyrotropin Regulation of Cholesterol Conversion Metabolism via the Akt-Dependent Pathway

Figure 6

Expression analysis of β-arrestin-mediated signal pathways related to cholesterol metabolism in 60 min TSH treatment. Cells were starved overnight and treated with 4 μM TSH. Total protein or nucleic protein was extracted at 0, 5, 30, and 60 min after TSH was added. Total protein was used for AKT phosphorylation level analysis, while nucleic component was used for the analysis of mature SREBP2, which is indicated as mSREBP2. (a) HepG2 cells, (b) ARRB1-knockdown HepG2-Cas9 cells, (c) ARRB2-knockdown HepG2 cells, and (d, e) grayscale analysis of relative phosphorylation level of AKT Thr308 and AKT Ser473 in TSH treatment of 0, 5, 30, and 60 min. (f) Grayscale analysis of relative expression level of mature SREBP2 in TSH treatment of 0, 5, 30, and 60 min. All experiments were repeated 3 times with similar results. indicated a statistical significance between genotypes. indicated a statistical significance between genotypes. (g) Differences of AKT phosphorylation and mature SREBP2 expression levels between ARRB1/2-knockdown and wild-type HepG2 cells treated with or without 60 min TSH treatment. (h, i, j) Grayscale analysis of AKT phosphorylation and mature SREBP2 expression levels between ARRB1/2-knockdown and wild-type HepG2 cells with or without TSH treatment for 60 min. indicated a statistical significance between time points within genotypes. indicated a statistical significance between time points within genotypes.

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