Fig. 3: mtHSP70 aggregation with LONP1 depletion. | Nature Communications

Fig. 3: mtHSP70 aggregation with LONP1 depletion.

From: LONP1 and mtHSP70 cooperate to promote mitochondrial protein folding

Fig. 3

a Rescue of mtHSP70 solubility with protease-deficient LONP1. Triton X-100 extraction and Western blotting were used to analyze protein solubility in LONP1 knockdown cells re-expressing doxycycline (DOX)-inducible RNAi-resistant LONP1WT and protease-deficient LONP1S855A. b Protein aggregation in LONP1 knockdown cells re-expressing ATPase-deficient LONP1. Triton X-100 extraction and Western blotting were used to examine protein solubility in LONP1 knockdown 143B cells re-expressing doxycycline (DOX)-inducible RNAi-resistant LONP1K529R. c Temporal features of mtHSP70 insolubility. mtHSP70 solubility was analyzed in 143B cells expressing DOX-inducible mtHSP70-HA. Cells were treated with DOX and 2 μM CDDO simultaneously or sequentially, as in Fig. 1f. d Comparison of OXA1L versus mtHSP70 solubility. 143B cells expressing DOX-inducible OXA1L-FLAG and mtHSP70-HA were treated with DOX and CDDO simultaneously or sequentially. Note the aggregation of mtHSP70 but not OXA1L in the sequential treatment. Scale bar, 10 μm. e Rescue of CDDO-induced mtHSP70 insolubility by CCCP or CHX. mtHSP70 solubility was analyzed in 143B cells expressing DOX-inducible mtHSP70-HA. Cells were treated with DOX and CDDO sequentially. CCCP or CHX was co-treated with CDDO as indicated.

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