Fig. 6. DELE1 responds to various types of perturbed mitochondrial protein import.
a 293T cells were transiently transfected with the specified constructs and DELE1 processing upon 4 h OM treatment was analyzed by immunoblotting; scHB, Saccharomyces cerevisiae heme-binding domain. scHB*, destabilized HB domain. b CHOPNeon fluorescence in the indicated cell lines was analyzed 20 h after transfection of mCherry fused to the sorting signals of COX5A or COX8A. Graph depicts mean ± s.d. of n = 3 independent experiments each containing three biological replicates. Statistical significance compared to WT within each construct is shown and was assessed using two-way ANOVA with Tukey’s multiple comparisons correction. COX5A-mCherry: ****P < 0.0001, ns ≥ 0.1712; COX8A-mCherry: ns ≥ 0.9994; mCherry: ns ≥ 0.3946. c 293T cells stably expressing the indicated DELE1-PINK1 chimeras were treated with DMSO or CCCP for 4 h and localization of the indicated HA-tagged proteins was analyzed by confocal microscopy. Scale bars, 10 μm. Nuclei (DAPI, blue), mitochondria (TRAP1, pink), HA (green). d, e CHOP induction upon knockdown of TIM23 or TOM40 by siRNA transfection into HeLa (d) or 293T (e) WT or DELE1 knockout (ΔDELE1) cells was analyzed by immunoblotting at the indicated times post-transfection.