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. 2022 Oct 11;10(6):2572–2585. doi: 10.1016/j.gendis.2022.10.003

Figure 4.

Figure 4

LiCl treatment restores Wnt signaling activity and proliferation of EMC1 KD HRECs. (A) Relative luciferase activity in Ctrl, EMC1 KD, and EMC1 KD HEK 293STF cells treated with LiCl. pGL4-Renilla plasmid was transfected as an internal control. Error bars, SDs. The P-values are from multiple comparisons in one-way ANOVA with Tukey's multiple comparison test (n = 6); ∗∗P < 0.01, ∗∗∗∗P < 0.0001. (B–G) Western blot and quantification analysis of the expression levels of CyclinD1, c-Myc, p-GSK3β, β-catenin, and FZD4 in Ctrl and EMC1 KD HRECs treated with LiCl or NaCl. Error bars, SDs. The P-values are from multiple comparisons in two-way ANOVA with Sidak's multiple comparison test (n = 3); ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, ∗∗∗∗P < 0.0001. ns, not significant. (H) Representative immunofluorescence images of EMC1 KD HRECs treated with NaCl or LiCl and labeled with EdU, β-catenin, or CyclinD1 and DAPI (blue). Scale bars, 20 μm (EdU and β-catenin) and 10 μm (CyclinD1). (I–K) Quantification of the percentage of EdU + nuclei (I), and relative levels of β-catenin (J) and CyclinD1 (K) in EMC1 KD HRECs treated with NaCl or LiCl. Error bars, SDs. Student's t-test (n ≥ 6), ∗∗∗∗P < 0.0001.