Additional file 1 of Fibroblast-to-cardiomyocyte lactate shuttle modulates hypertensive cardiac remodelling
Creators: name: Wei, Tong
name: Guo, Yuetong
name: Huang, Chenglin
name: Sun, Mengwei
name: Zhou, Bin
name: Gao, Jing
name: Shen, Weili
Publisher: name: figshare
Year: 2024
Subjects: subject: Biochemistry
subject: Space Science
subject: Cell Biology
subject: Genetics
subject: FOS: Biological sciences
subject: Molecular Biology
subject: Physiology
subject: Pharmacology
subject: Biotechnology
subject: Environmental Sciences not elsewhere classified
subject: Biological Sciences not elsewhere classified
subject: Cancer
subject: Computational Biology

description: Additional file 1: Fig. S1. A and B WT and GCN5L1CKO mice were subjected to saline or Ang II for 4 weeks. Representative immunoblot images showing GCN5L1 and Postn protein expression in cardiac myofibroblasts isolated from WT or GCN5L1CKO mice (A). The systolic blood pressure of WT and GCN5L1CKO mice with saline or Ang II infusion (B). The data are shown as the mean ± SEM (n=8/group). P values were calculated by one-way ANOVA.**p &lt;0.01 vs. WT-saline. Fig. S2. A Immunofluorescence images of CFs stained with 2-NBDG (green) and Hoechst (blue). Scale bar=100 μm. B and C Representative immunoblot images showing Glut1 protein expression in CFs treated with or without Ang II after infection with an empty vector or sh-GCN5L1 (B). The quantification of these proteins (C). n≥2/group. D and E Representative immunoblot images showing Glut1 protein levels in CFs treated with or without Ang II after infection with an empty vector or LV-GCN5L1 (D). The quantification of these proteins (E). n≥2/group. F and G, Representative immunoblot images showing α-SMA and vimentin protein expression in CFs pre-treatment with 2-DG then cultured with or without Ang II (F). The quantification of these proteins (G). n=5/group. H and I Representative images of the CF wound assay. Scale bar=200 μm (H). The percentage of the wound closed (I). n=13/group. J and K Representative immunoblot images showing α-SMA and vimentin protein expression in CFs pre-treated with UK5099 for 1h then cultured with or without Ang II (J). The quantification of these proteins (K). n≥3/group. L and M Representative immunoblot images showing α-SMA and vimentin protein expression in CFs were pre-treated with methyl pyruvate for 1h then cultured with or without Ang II. (L). The quantification of these proteins (M). n≥3/group. The data are shown as the mean ± SEM. P values were calculated by one-way ANOVA. *p&lt;0.05,**p&lt;0.01. Fig. S3. A and B Immunofluorescence of Ac-MPC2K19 (red) and DAPI (blue) in Ang II treated CFs after transfection with an empty vector, sh-GCN5L1 or LV-GCN5L1. Scale bar = 100 μm (A). Quantification of the relative fluorescence intensity (B). n = 10/group. C, Schematic diagram showing isotope-tracing experiments. D, CFs were transfected with Flag-MPC2WT, Flag-MPC2K19Qor Flag-MPC2K19R. [U-13C3]-pyruvate was added into culture media for 30 min. Then, tracing analysis from [U-13C3]-pyruvate was performed. Intracellular abundance of citrate (M+2), α-KG (M+2), succinate (M+2), fumarate (M+2), Malate (M+2) was calculated. n=3/group. The data are shown as the mean ± SEM. P values were calculated by one-way ANOVA. *p &lt;0.05, **p &lt;0.01. Fig. S4. A Representative immunoblot images showing MCT1 protein expression in cardiomyocytes after infection with an empty vector or sh-MCT1. B Quantification of the MCT1 level. n=6/group. The data are shown as the mean ± SEM. P values were calculated by t-test. **p &lt;0.01 vs. vector. C Representative immunoblot images showing MCT1 protein expression in the hearts of WT and MCT1CKO mice. D Quantification of the MCT1 level. n = 4/group. P values were calculated by t-test. **p &lt;0.01 vs. WT mice.