Extended Data Fig. 4: Characterization of PEX39 loss in human cells.
From: PEX39 facilitates the peroxisomal import of PTS2-containing proteins
a, Quantification of mature ACAA1, mature AGPS, PEX7, and PEX5 in HsPEX39-knockout cells using band intensities of immunoblots prepared per Fig. 2f. Loading-control intensities were used to normalize protein abundance across samples and the normalized abundances were then divided by the mean value of the corresponding control cells. The names of quantified proteins are shown on the bottom. Precursor forms of ACAA1 and AGPS were undetectable and thus not quantified. Data are mean ± standard error of the mean (SEM) (n = 3), and P values were calculated using unpaired, two-tailed t-tests. b, Immunoblot analysis of cells generated via lentiviral transduction of Cas9 and multiple independent sgRNAs against HsPEX39 (sgHsPEX39_1-3) or a negative control sgRNA against the AAVS1 gene (sgAAVS1) in the CAKI-2 and NCI-H1792 cell lines. Note that the residual HsPEX39 signal in these cells is because not all cells in the population will have complete loss of the protein. Solid and open red arrowheads indicate mature and precursor forms, respectively. Precursor forms of ACAA1 and AGPS were undetectable. CANX is a loading control. Short and long exposures are denoted s.e. and l.e., respectively. c, Immunoblots of whole-cell (Cell), cytosolic (Cyto), and organellar (Org) fractions prepared from the CAKI-2 cells expressing sgAAVS1 (negative control) or sgHsPEX39_1 described in b. Solid and open red arrowheads indicate mature and precursor forms, respectively. The cellular fractions that RPS6KB1 and CANX are markers for are indicated in parentheses.
