Journal: Advanced Science

Article Title: PRDM16 Reduces Cellular Senescence by Upregulating GSTM1

doi: 10.1002/advs.202501233

Figure Lengend Snippet: PRDM16 inhibited oxidative DNA damage and improved glutathione metabolism. a) Diagram detailing combined analysis of kidney scRNA‐seq data from 9‐month‐old WT and Prdm16 KO mice (n = 3) and bulk RNA sequencing (bulk RNA‐seq) data from irradiated PRDM16‐overexpressing and control HK‐2 cells (n = 3). Gene Ontology (GO) enrichment analysis was performed to differentially expressed genes of proximal convoluted tubule (PCT) and distal convoluted tubule (DCT) from scRNA‐seq, as well as differentially expressed genes of HK‐2 cells from RNA‐seq. Common GO terms were identified by intersection. b) Representative IF images of 8‐oxo‐dG in the kidney. The ratio of 8‐oxo‐dG positive cells to total cells was calculated (n = 5 for Prdm16 fl/fl group, n = 6 for the other 3 groups). Scale bar: 20 µm. c) Representative IF images of 8‐oxo‐dG and Dihydroethidium (DHE) in HK‐2 cells. The percentage of 8‐oxo‐dG positive cells in total cells and relative fold change of DHE intensity were calculated. Scale bar: 20 µm. d) Enrichment plot from the GSEA results based on scRNA‐seq data of PCT. e) Diagram detailing the enzymes involved in glutathione metabolism. qPCR analysis of Gclc , Gclm and Gss in the kidney of aging mice treated with negative control lentivirus (NC‐LV) or PRDM16 overexpression lentivirus (PR‐LV). GSH/GSSG ratio and GST activity of irradiated HK‐2 cells and the kidney of aging mice were detected and calculated. (n = 3 for HK‐2 cells and n = 6 for mice). Data are mean ± SEM. * p < 0.05, ** p < 0.01 and *** p < 0.001. n.s: not significant. One‐way ANOVA followed by Tukey's post‐test (b, c and e).

Article Snippet: The GST activity of kidney tissues or HK‐2 cells were measured using GST Activity Assay Kit (E‐BC‐K278‐S, Elabscience) according to the instruction.

Techniques: RNA Sequencing, Irradiation, Control, Negative Control, Over Expression, Activity Assay

Journal: Advanced Science

Article Title: PRDM16 Reduces Cellular Senescence by Upregulating GSTM1

doi: 10.1002/advs.202501233

Figure Lengend Snippet: PRDM16 inhibited oxidative DNA damage and improved glutathione metabolism. a) Diagram detailing combined analysis of kidney scRNA‐seq data from 9‐month‐old WT and Prdm16 KO mice (n = 3) and bulk RNA sequencing (bulk RNA‐seq) data from irradiated PRDM16‐overexpressing and control HK‐2 cells (n = 3). Gene Ontology (GO) enrichment analysis was performed to differentially expressed genes of proximal convoluted tubule (PCT) and distal convoluted tubule (DCT) from scRNA‐seq, as well as differentially expressed genes of HK‐2 cells from RNA‐seq. Common GO terms were identified by intersection. b) Representative IF images of 8‐oxo‐dG in the kidney. The ratio of 8‐oxo‐dG positive cells to total cells was calculated (n = 5 for Prdm16 fl/fl group, n = 6 for the other 3 groups). Scale bar: 20 µm. c) Representative IF images of 8‐oxo‐dG and Dihydroethidium (DHE) in HK‐2 cells. The percentage of 8‐oxo‐dG positive cells in total cells and relative fold change of DHE intensity were calculated. Scale bar: 20 µm. d) Enrichment plot from the GSEA results based on scRNA‐seq data of PCT. e) Diagram detailing the enzymes involved in glutathione metabolism. qPCR analysis of Gclc , Gclm and Gss in the kidney of aging mice treated with negative control lentivirus (NC‐LV) or PRDM16 overexpression lentivirus (PR‐LV). GSH/GSSG ratio and GST activity of irradiated HK‐2 cells and the kidney of aging mice were detected and calculated. (n = 3 for HK‐2 cells and n = 6 for mice). Data are mean ± SEM. * p < 0.05, ** p < 0.01 and *** p < 0.001. n.s: not significant. One‐way ANOVA followed by Tukey's post‐test (b, c and e).

Article Snippet: The GST activity of kidney tissues or HK‐2 cells were measured using GST Activity Assay Kit (E‐BC‐K278‐S, Elabscience) according to the instruction.

Techniques: RNA Sequencing, Irradiation, Control, Negative Control, Over Expression, Activity Assay