Fig. 4

Fkbp51 KO mice have a higher expression of Parkin than WT. A Quantitative PCR analyses reveal that Fkbp51 KO liver expresses more Park2 mRNA relative to WT in both control and CCl₄-treated conditions. B Immunohistochemical labeling demonstrates an increase in Parkin in KO, and a CCl₄–associated increase in both WT and KO. C Quantitation of AOD of Parkin in liver sections. D Western blotting confirms a higher level of Parkin in control (Con) KO liver, with an apparent increase of Parkin in WT following CCl₄ treatment. E–H Increased Parkin expression in total KO liver lysates as well as in enriched mitochondrial and cytoplasmic fractions as compared to WT. Graphs represent mean values ± SEM from 6 mice for each group. p values were determined by student’s unpaired t-test (F–H) or two-way ANOVA (A, C, D) with the statistical significance labeled as follows: *as p < 0.05, **as p < 0.01, *** as p < 0.001 and **** as p < 0.0001. I A comparison of Fkbp51 KO and WT MEFs confirms that the absence of FKBP51 is associated with higher levels of Parkin, more intense MitoTracker signal, and localization to the mitochondria. Magnified images outlined by the yellow boxes in the merged images are presented on right. Immunofluorescent labeling of Parkin and MitoTracker showed higher intensity of Parkin signal in KO than WT MEF cells, particularly in the perinuclear area and along the mitochondrial network. J Immunofluorescent labeling reveals the co-localization of FKBP51 and Parkin with mitochondria in HepG2 cells transfected with Flag-Fkbp51. Key: WT, wild type; KO, Fkbp51 KO; AOD, average optical density; Con, control; Mito, mitochondria