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. 2020 Apr 19;10(8):1426–1439. doi: 10.1016/j.apsb.2020.03.008

Figure 4.

Figure 4

S-Palmitoylation facilitates an accurate intracellular localization and cell proliferation for CLDN3. (A) Confocal images show subcellular localization of endogenous ZO-1, FLAG-tagged CLDN3 WT or mutants in A2780 cell line. Cells expressing FLAG-tagged CLDN3 3CS were treated with MG132 for 4 h before fixed and immunofluorescence staining with indicated antibodies. The y–z axis shows the location of ZO-1 and CLDN3 in basal and apical regions. (B) Quantitative analysis of correlated co-localization of CLDN3 (green) and ZO-1 (red) in (A), representing by the mean of Manders' coefficient with threshold (tM1 and tM2) (n = 7 for each group; ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001). (C) Validation of the expression of FLAG-tagged CLDN3 WT and mutants in shCLDN3 OVCAR8 cells by immunoblotting. (D) Representative data from soft agar colony formation of shCLDN3 OVCAR8 cells expressing FLAG-tagged CLDN3 WT or mutants. 3.0 × 103 cells were cultured in RPMI-1640 medium contained 10% FBS and 0.35% agarose gel for 21 days. Cell colonies were stained by MTT. (E) The quantification of the colony numbers in soft agar colony formation assay (D). Values with error bars indicate mean ± SD of three independent replicates (∗P < 0.05; ∗∗P < 0.01).