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. 2021 Oct 23;10(2):1994351. doi: 10.1080/21688370.2021.1994351

Figure 4.

Figure 4.

In the liver of OZ rats with steatosis, TJP2 mRNA and ZO-2 protein diminish while YAP concentrates at the nucleus of hepatocytes. A) Frozen sections from livers of lean (LZ) and obese Zucker rats (OZ) of 12 weeks of age were processed for immunofluorescence with antibodies against ZO-1 and ZO-2. Nuclei were stained with DAPI. Bar, 10 μm. Arrows indicate TJ staining as a pair of dots; arrowheads show TJ staining at parallel strands along intrahepatic bile ducts. Left panel, representative images; right panel, quantitative analysis. Statistical analysis was done with GraphPad Prism 5, Mann Whitney test, ***p < .001. B) Western blot of ZO-2 present in a liver lysate derived from LZ and OZ rats. Upper panel, representative Western blot; lower panel, quantitative analysis of three independent experiments. Results are mean ± SEM. Statistical analysis was done with the student’s t-test. *p = .03. C) qRT-PCR amplification of TJP2 mRNA from LZ and OZ rats. Data of LZ rat normalized to 1. Statistical analysis was done with GraphPad Prism 5, Student’s t-test, *p < .05. Results were obtained from three independent experiments. Data were normalized against the geometric mean of actin mRNA. D) Frozen sections from livers of LZ and OZ of 12 weeks of age were processed for immunofluorescence with antibodies against Sp1. Nuclei were stained with DAPI. Bar, 10 μm. Upper panel, representative images; lower panel, quantitation of immunofluorescence for nuclear Sp1. The nuclei of 200 cells per condition derived from three independent experiments were quantitated. Statistical analysis was done with Mann Whitney test, ****p < .0001. E) Frozen sections from livers of LZ and OZ of 12 weeks of age were processed for immunofluorescence with antibodies against YAP. Nuclei were stained with DAPI. Bar, 10 μm. Upper panel, representative images; lower panel, quantitation of immunofluorescence for nuclear YAP. The nuclei of 100 cells per condition derived from three independent experiments were quantitated. Statistical analysis was done with Mann Whitney test, ****p < .0001.