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. 2020 Oct 17;11(10):873. doi: 10.1038/s41419-020-03081-w

Fig. 2. RNF40 loss impairs oncogenic properties of HER2-BC cells in vitro.

Fig. 2

A Western blot validation of RNF40 knockdown efficiency and decreased H2Bub1 levels in HCC1954 cells. B Representative bright-field pictures of control and RNF40 siRNA-transfected HCC1954 and SKBR3 cells. Scale bars (white): 500 µm. C, D Proliferation curves (C) and clonogenic assays (D) of control and RNF40-depleted HCC1954 and SKBR3 cells. Quantification of the occupied area in clonogenic assays is shown for both cell lines (D, lower panel). Student t-test. E Tumorsphere formation assay of control and RNF40-depleted HCC1954 cells (left panel). Quantification of the tumorsphere number normalized to the control condition (right panel). Student t-test. F Representative pictures from immunofluorescence detection of RNF40 and the proliferation marker Ki67 in control and RNF40-depleted HCC1954 and SKBR3 cells (upper panel). Scale bars = 60 μm. Quantification of the Ki67 immunofluorescence intensity of single nuclei in control and RNF40-depleted HCC1954 and SKBR3 cells (lower panel). The median intensity values of the respective groups are provided as green bars. Mann–Whitney test. G Transwell migration assay of control and RNF40-depleted HCC1954 cells with representative results (left panel) and the corresponding quantification (right panel). *p < 0.05, **p < 0.01, ***p < 0.005. All experiments were performed in biological triplicates. Error bars: SEM.