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Fig. 3

ID
ZDB-IMAGE-211018-45
Source
Figures for Yang et al., 2021
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Figure Caption

Fig. 3 Functional impairment of <italic>GBP1</italic> variants identified in congenital hypothyroidism (CH) patients.

(a) The rescuing effects of co-expression of wild-type (WT) hGBP1 or mutated hGBP1 messenger RNA (mRNA) (hGBP1-H150Y or hGBP1-L187P) on thyroid development are displayed. Bars = 50 μm. (bd) Statistical analysis of the thyroid area (b), angle between two posterior strands (c), and length to width ratio (d) in WT and gbp1 morphants with WT or mutated hGBP1 mRNA overexpression. N = 12. (e) Rescuing effects tshba expression in zfgbp1 morphants with WT or mutated hGBP1 mRNA overexpression. Means ± SEM are shown for three independent experiments. (f) The expression of endogenous hGBP1 and β-catenin in TPC1 cells was detected by immunofluorescence. Bars = 20 μm. The enlarged image shows that hGBP1 colocalized with β-catenin in the cytoplasm of TPC1 cells. (g) Co-immunoprecipitation (Co-IP) assessment of the interaction between WT and mutated hGBP1 (p.H150Y, p.L187P) with β-catenin. (h) Co-IP assessment of the interaction between the truncated hGBP1 (p.E336fs) with β-catenin. (i) Western blot examination of cytosol and membrane β-catenin content with overexpression of N-terminal flag fused hGBP1. Na/K+ ATPase was used as a loading control for the membrane and α-tubulin for the cytosol fraction. (j) The effect of GBP1 knockdown in TPC1 cells on the content of cytosol and membrane β-catenin. Na/K+ ATPase was used as a loading control for the membrane and α-tubulin for the cytosol fraction. (k) The effect of introducing N-terminal flag fused GBP1 on the formation of adhesion complex in TPC1 cells. (l) Restoring effects of WT or mutated hGBP1 overexpression on the subcellular β-catenin levels in GBP1-deficient TPC1 cells. Na/K+ ATPase was used as a loading control for the membrane and α-tubulin for the cytosol fraction. (m) Rescuing effects of WT or mutated hGBP1 overexpression on cellular adhesion complex formation in GBP1-deficient TPC1 cells. Means ± SEM are shown for three independent experiments. n.s. not significant; **P < 0.01; ***P < 0.001 (Student’s t-test).

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