Figure 4.

Peroxisome-derived H₂O₂ does not trigger pexophagy in po-mKeima/DD-DAO Flp-In T-REx 293 cells. DD-DAO Flp-In T-REx 293 cells stably expressing po-mKeima were cultured as described in the legend to Figure 3. The cells were subsequently transferred to different assay media (DPBS, mDMEM, or mMEMα; for details, see Section 2) supplemented with 10 mM L- or D-Ala. (A) Representative fluorescence overlay images of po-mKeima on excitation at 440 (false color: green) and 586 (false color: red) nm. Scale bar, 10 µm. (B) The po-mKeima 586/440 nm excitation fluorescence ratios. The data represent the values of the independent biological replicates. The corresponding D- and L-Ala data were statistically compared, but no significant differences were found. (C) Examples of flow cytometry plots for cells transferred to DPBS for 2 h. The different colors represent the cell density at a given position. (D) Quantification of the percentage of cells in the gated area. Data are shown as the mean ± SD (n = 3). The corresponding D- and L-Ala data were statistically compared (ns, non-significant; **, p < 0.01).