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. 2024 Sep 26;25(19):10369. doi: 10.3390/ijms251910369

Figure 3.

Figure 3

NPHP3 overexpression in HeLa cells reduced the death. (A–C) HeLa cells were transfected with pEGFP-C1 or pEGFP-HPHP3 for 36 h. Cell lysates were prepared and western blot analysis was performed to detect the expression of various proteins, NPHP3-GFP (A top), NPHP3 (A top and middle), GFP (A middle) and tubulin (A bottom). Cells in each group were treated with 1 nM VBL in the presence of FBS for 24 h. Then, cells were fixed and permeabilized and the nucleus was stained with DAPI (blue). Apoptotic cell death out of GFP-positive cells was observed at 400× magnification with 40× objective under the fluorescence microscope (B,C). The image with GFP fluorescence is representative of ≈30 pictures. White arrows indicated the dying cells with nuclear condensation (B). Bar graph represents the relative cell death (C). Processing (such as changing brightness and contrast) is applied equally to control across the entire image (A,B). Data in a bar graph represents the means ±SEM. ** p < 0.01; significantly different from the VBL-untreated control group. && p < 0.01; significantly different from the VBL-treated group (C).