Additional file 1 of Elevated CHCHD4 orchestrates mitochondrial oxidative phosphorylation to disturb hypoxic pulmonary hypertension
Creators: name: Wang, Yu
name: Zeng, Zhenyu
name: Zeng, Zhaoxiang
name: Chu, Guojun
name: Shan, Xinghua
Publisher: name: figshare
Year: 2024
Subjects: subject: Biophysics
subject: Biochemistry
subject: Medicine
subject: Cell Biology
subject: Molecular Biology
subject: Physiology
subject: FOS: Biological sciences
subject: Chemical Sciences not elsewhere classified
subject: Ecology
subject: Immunology
subject: FOS: Clinical medicine
subject: Biological Sciences not elsewhere classified
subject: Marine Biology
subject: Cancer

description: Additional file 1: Figure S1. CHCHD4 is identified as a regulator of PAH. Related to Fig. 1. A Quantification of ratio of pulmonary arterial medial thickness to total vessel size (media/CSA) in indicated group (n = 6). Related to Fig. 1D. B Quantification of immunoblotting of CHCHD4 (n = 6). Related to Fig. 1C. C Quantitative analysis of α-SMA and CHCHD4 fluorescence intensity by Image J software (n=6). Related to Fig. 1E. D Representative immunofluorescence of Vimentin (green), CHCHD4 (red) and DAPI (blue) in lung tissues from indicated group. And quantitative analysis of CHCHD4 fluorescence intensity by Image J software (n = 6). Data are shown as the mean ± SEM. P value is showed in each figure. Figure S2. Protocols. A Experimental schedule of SD rats received AAV1-Chchd4 or CTR injection. Representative immunoblotting of CHCHD4 in lung tissues from animals with AAV1-CTR or Chchd4. Related to Fig. 3. B VSMCs, ECs and other cells were isolated from SD rats. Related to Fig. 1. C Experimental schedule of primary PASMCs received Chchd4 overexpression or knockdown in vitro. Related to Fig. 4. Figure S3. CHCHD4 affects hypoxia-induced migration in PASMCs. A The percent of wound closure of PASMCs received len-CTR or len-CHCHD4 transduction was counted (n = 6 fields from 3 independent experiments). Related to Fig. 4B. B The percent of wound closure of PASMCs received siCTR or siCHCHD4 transduction was counted (n = 6 fields from 3 independent experiments). Related to Fig. 4G. Data are shown as the mean ± SEM. P value is showed in each figure. Figure S4. Overexpression of CHCHD4 improves hypoxia-induced mitochondrial dysfunction. Related to Fig. 5. A Quantification of basal respiration, ATP production–coupled respiration, maximal respiration, and spare respiratory capacity from oxygen consumption rate (OCR) in PASMCs. Related to Fig. 5G. B Quantification of glycolysis, glycolytic capacity and glycolytic reserve from extracellular acidification rate (ECAR) in PASMCs. Related to Fig. 5H. Data are shown as the mean ± SEM. P value is showed in each figure. Figure S5. CHCHD4 modulates mitochondrial dynamics. Related to Fig. 6. A. Quantification of immunoblotting of Opa1, Drp1 and Mfn1 in lung PA tissues from indicated groups (n = 6). Related to Fig. 6C. B Quantification of immunoblotting of Opa1, Drp1 and Mfn1 in isolated PASMCs from indicated groups (n = 6). Related to Fig. 6D. Data are shown as the mean ± SEM. P value is showed in each figure. Figure S6. SAM50 knockdown abolishes the protective effects of CHHCD4 during hypoxia. Related to Fig. 7. Quantification of immunoblotting of CHCHD4, SAM50, Opa1, Drp1 and Mfn1 in isolated PASMCs from indicated groups (n = 6). Related to Fig. 7C.