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. 2016 Aug 3;12(10):1759–1775. doi: 10.1080/15548627.2016.1199300

Figure 5.

Figure 5.

Autophagic cargo in WT and bs MEFs. Immunostaining of WT (A) and bs (B) MEFs for ubiquitin. Immunoblotting of WT and bs cell lysates (C) confirmed accumulation of ubiquitinated proteins in bs MEFs; immunoblotting for ACTB served as a loading control. Immunostaining of WT (D) and bs (E) MEFs for ABCD3 as a peroxisomal marker; quantification of ABCD3-positive puncta (F) confirmed a significant (P = 0.0001; n = 30) accumulation of ABCD3 in bs MEFs. Immunostaining of WT (G) and bs (H) MEFs for COX4I1 as a mitochondrial marker. Western blotting of WT and bs lysates for TOMM20 as a mitochondrial marker revealed in bs lysates significantly greater (P = 0.0063; n = 3) levels of TOMM20 relative to ACTB (I). Staining of WT (J) and bs (K) MEFs with the mitochondrial dye MitoTracker Red established that accumulating mitochondria in bs MEFs remained polarized. Nuclei in all images were stained with DAPI. Scale bar: 5 μm. The data in (F) and (I) are presented as the means ± SEM and significance was established with the Student t test.