Figure 2.

MCT2 overexpression increases peroxisome surface, decreases peroxisome number, and does not affect the expression of key proteins involved in β-oxidation in 22Rv1 PCa cells: (A) peroxisomes form clusters in 22Rv1 cells, as shown by (a) immunofluorescence analysis of PEX14 by confocal microscopy and (b) DAB-staining based transmission electron microscopy; (P) peroxisomes, (M) mitochondria, and (N) nucleus. Nuclei are shown in blue (stained with Hoechst 33258) in (a). White bar represents 5 μm and black bar represents 0.5 μm. (B–E) Analysis of modifications in peroxisome morphology and number in 22Rv1 cells transfected with Myc-MCT2, when compared with nontransfected cells. (B) Immunofluorescence analysis of (a) Myc and (b) PMP70 by confocal microscopy in Myc-MCT2-transfected 22Rv1 cells; (c) merge image of a and b, presenting nuclei in blue. (C) Quantification analysis of alterations in peroxisomes’ area (pixel²), presented as the mean of peroxisome area per total cell area. (D) An example of a representative single-cell analysis is presented as the mean of peroxisome area in a cell. (E) Quantification analysis of changes in peroxisomes’ number, presented as the mean of peroxisome number per cell area (pixel²). Data represent means of three independent experiments and the bars represent SEM of the mean. * p < 0.05 **** p < 0.0001. (F) Western blot analysis showing the expression levels of MCT2, CAT, ACOX3, PMP70, and TUB in control and Myc-MCT2-transfected 22Rv1 cells. A densitometric quantification of the immunoblots is presented in Figure S1.