Fig. 5. CdCl₂ induces RB expression and its mitochondrial translocation both in vivo and in vitro.

a, b The ICR mice were intraperitoneally injected with CdCl₂ (1 mg/kg bw, n = 3) or physiological saline (Ctrl group, n = 3) for 7 days. a Liver slides were subjected to IHC using anti-RB and -Drp1 antibodies, respectively. The scale bar is 50 μm. b Whole cell lysates and mitochondrial fraction of liver tissues were prepared for Western blot with the indicated antibodies. c Western blot was applied to detect the expression levels of RB and Drp1 after CdCl₂ (10, 20, and 40 μM) exposure to L02 cells for 6 h. (arrow denotes the specific band.) d Cytoplasmic (Cyto) and mitochondrial (Mito) fractions of L02 cells were prepared for Western blot to determine the expression of proteins as indicated. e–h L02 cells were treated with CdCl₂ (20 μM) for 6 h. e Cells were subjected to IF staining with MitoTracker (red), anti-RB antibody (green), and DAPI (blue, staining nuclei). The scale bar is 10 μm. f Mito/RB signal overlap was analyzed with Pearson’s correlation. Red-green overlap sections were quantified by using ImagePro-plus 6.0. Ten fields of view were calculated for each group. g The images of L02 cells were labeled with MitoTracker (red) and immunostained with antibodies of anti-Drp1 (blue) and anti-RB (green). The scale bar is 10 μm. h Pearson’s correlation for the co-localization of Mito-RB, Drp1-RB, and Mito-Drp1 was shown in bar graphs quantified by using ImagePro-plus 6.0. Ten fields of view were calculated for each group. Data are expressed as the mean ± standard deviation (SD). P < 0.05, *significantly different from Ctrl