Figure 3. Peroxisomal membrane targeting of Pex3 is essential for the induction of pexophagy. (A) Schematic representation and alignment of N-terminal amino acid sequence of the PEX3-HA₂ variants, including Mito-PEX3-HA₂ and Cyto-PEX3-HA₂. The 39 N-terminal amino acid peroxisome-targeting sequences were replaced with the 69 N-terminal amino acids of TOMM20 (Mito-PEX3) or deleted (Cyto-PEX3). (B) CHO-K1 cells were transfected with empty vector or PEX3-HA₂ variants. After 12 h, the cells were fixed and immunostained with antibodies against HA (b, g, l, and q), TOMM20 (c, h, m, and r) and PEX14 (d, i, n, and s). Merged views of the PEX3-HA₂ variants and TOMM20 are shown in (f, k, and p). (C) The percentage of cells exhibiting pexophagy was calculated as in Figure 1B. Data are presented as the mean ± SD of 3 replicates.