Figure 2 : Heterologous expression of human ClpP complements the longevity phenotype of ΔPaClpP.

From: Human CLPP reverts the longevity phenotype of a fungal ClpP deletion strain

Figure 2

(a) Representative western blot analysis of mitochondrial protein extracts from wild-type, ΔPaClpP and ΔPaClpP/HsClpP_OEx1. The HsCLPP-specific antibody detects the ~26 kDa HsCLPP monomer only in the ΔPaClpP/HsClpP_OEx1 sample. PaPORIN (PaPOR) was detected as a loading control. (b) Western blot analysis of the same mitochondrial protein extracts as in (a) following BN-PAGE. Incubation of the western blot membrane with a PaCLPP-specific antibody visualized two oligomers with apparent molecular masses of ~170 and 340 kDa in the wild-type sample. In the ΔPaClpP/HsClpP_OEx1 sample, two distinct oligomers with apparent molecular masses of ~220 and 440 kDa were detected after incubation with an HsCLPP-specific antibody. Additionally, the ΔPaClpP/HsClpP_OEx1 (den.) sample was heat denatured for 10 min at 95 °C, after which the HsCLPP oligomers could not be detected anymore. (c) Lifespan of wild-type (23.3±0.4; n=39), ΔPaClpP (39.8±1.9; n=37; P=7.2E-13), ΔPaClpP/HsClpP_OEx1 (26.2±0.7; n=40; P=6.8E-04) and ΔPaClpP/HsClpP_OEx2 (26.2±0.7; n=39; P=2.5E-03) isolates at 27 °C. Data given in parentheses are mean lifespan±s.e. in days. P-values were determined in comparison with the wild-type sample by two-tailed Wilcoxon rank-sum test.