Extended Data Fig. 7: MIEF1 phosphorylation mediates mitochondrial mechanotransduction.
a, Representative pictures of SREBP2 localization in MCF10A-RAS cells. Scale bar 10 μm. Refers to Fig. 8a. b, Representative pictures of YAP localization in MCF10A-RAS cells. Scale bar 10 μm. Refers to Fig. 8b. c, SREBP2 localization in MCF10A-RAS cells reconstituted as in Fig. 8a, but with an independent set of MIEF1-2 siRNAs (siMIEF1/2b) (siCO DMSO and siCO YM as in Fig. 8a, n = 57 (siMIEF1/2b DMSO), n = 65 (siMIEF1/2b YM), n = 56 (siMIEF1/2b WT low level YM), n = 61 (WT medium level), n = 58 (WT high level), n = 54 (SA low level), n = 66 (SA medium level), n = 60 (SA high level), n = 62 (SE low level), n = 64 (SE medium level) and n = 58 (SE high level) from three experiments). d, YAP localization in MCF10A-RAS cells reconstituted as in Fig. 8b, but with an independent set of MIEF1-2 siRNAs (siMIEF1/2b) (siCO DMSO and siCO YM as in Fig. 8b, n = 77 (siMIEF1/2b DMSO), n = 79 (siMIEF1/2b YM), n = 81 (siMIEF1/2b WT low level YM), n = 69 (WT medium level), n = 72 (WT high level), n = 73 (SA low level), n = 78 (SA medium level), n = 81 (SA high level), n = 84 (SE low level), n = 89 (SE medium level) and n = 88 (SE high level) from three experiments). e, Representative pictures of SREBP2 localization in MCF10A-RAS cells. Scale bar 10 μm. Refers to Fig. 8c. f, Representative pictures of YAP localization in MCF10A-RAS cells. Scale bar 10 μm. Refers to Fig. 8d. g, SREBP2 localization in Mief1-Mief2-Mff knockout MEFs transiently transfected to reconstitute different levels of the wild-type (WT), phosphorylation-mimicking (SE) and phosphorylation-null (SA) MIEF1 variants. Where indicated, cells were treated with ROCK-MLCK inhibitors (YM) (n = 71 cells (DMSO), n = 65 (YM), n = 67 (WT low level), n = 67 (WT medium level), n = 67 (WT high level), n = 63 (SA low level), n = 68 (SA medium level), n = 69 (SA high level), n = 68 (SE low level), n = 71 (SE medium level), n = 65 (SE high level) from three experiments). h, YAP localization in Mief1-Mief2-Mff knockout MEFs transiently transfected to reconstitute different levels of the wild-type (WT), phosphorylation-mimicking (SE) and phosphorylation-null (SA) MIEF1 variants. Where indicated, cells were treated with ROCK-MLCK inhibitors (YM) (n = 92 cells (DMSO), n = 89 (YM), n = 99 (WT low level), n = 97 (WT medium level), n = 102 (WT high level), n = 121 (SA low level), n = 99 (SA medium level), n = 105 (SA high level), n = 97 (SE low level), n = 92 (SE medium level), n = 104 (SE high level) from three experiments). Data are presented as mean values ± s.d. p-values were calculated based on two-tailed Student’s t-tests. In e,g, this was calculated on nuclear SREBP2. In f,h,this was calculated on nuclear YAP. Source numerical data are available in the source data.
