Journal: Scientific Reports

Article Title: Quercetin-4’-O-β-D-glucopyranoside inhibits ferroptosis though SIRT5-mediated desuccinylation of TFR1 in diabetic nephropathy

doi: 10.1038/s41598-026-41148-4

Figure Lengend Snippet: QODG inhibited HG-induced ferroptosis in MPC5 cells. ( A) , CCK-8 was performed to analyze the cell ability of each group ( N = 3); ( B) , Flow cytometry was performed to analyze the relative fluorescence intensity of lipid peroxidation in each group ( N = 3); Cellular ( C) , SOD, ( D) , CAT, ( E) , MDA, and ( F) , iron contents in each group were determined by commercial kits ( N = 3); ( G) , Western blot was performed to assess the protein levels of GPX4, SLC7A11, and TFR1 in each group ( N = 3). ** p < 0.01.

Article Snippet: MPC5 cells were transfected with SIRT5-targeting short hairpin (sh) RNA (shSIRT5), shRNA negative control (shNC), pcDNA3.1 empty vector, or transferrin receptor 1 (TFR1) overexpression plasmid (pcDNA3.1-TFR1) using Lipofectamine 3000 (#L3000001; Thermo Fisher Scientific, Waltham, MA, USA) following the manufacturer’s protocol.

Techniques: CCK-8 Assay, Flow Cytometry, Fluorescence, Western Blot

Journal: Scientific Reports

Article Title: Quercetin-4’-O-β-D-glucopyranoside inhibits ferroptosis though SIRT5-mediated desuccinylation of TFR1 in diabetic nephropathy

doi: 10.1038/s41598-026-41148-4

Figure Lengend Snippet: Silence of SIRT5 promoted ferroptosis in HG-induced MPC5 cells. ( A) , The mRNA levels of SIRT5 after transfection of shSIR5 vector into MPC5 cells were analyzed by RT-qPCR ( N = 3); ( B) , CCK-8 was performed to analyze the cell ability of each group ( N = 3); ( C) , Flow cytometry was performed to analyze the relative fluorescence intensity of lipid peroxidation in each group ( N = 3); Cellular ( D) , SOD, ( E) , CAT, ( F) , MDA, and ( G) , iron contents in each group were determined by commercial kits ( N = 3); ( H) , Western blot was performed to assess the protein levels of SIRT5, GPX4, SLC7A11, and TFR1 in each group ( N = 3). ** p < 0.01.

Article Snippet: MPC5 cells were transfected with SIRT5-targeting short hairpin (sh) RNA (shSIRT5), shRNA negative control (shNC), pcDNA3.1 empty vector, or transferrin receptor 1 (TFR1) overexpression plasmid (pcDNA3.1-TFR1) using Lipofectamine 3000 (#L3000001; Thermo Fisher Scientific, Waltham, MA, USA) following the manufacturer’s protocol.

Techniques: Transfection, Plasmid Preparation, Quantitative RT-PCR, CCK-8 Assay, Flow Cytometry, Fluorescence, Western Blot

Journal: Scientific Reports

Article Title: Quercetin-4’-O-β-D-glucopyranoside inhibits ferroptosis though SIRT5-mediated desuccinylation of TFR1 in diabetic nephropathy

doi: 10.1038/s41598-026-41148-4

Figure Lengend Snippet: Silence of SIRT5 enhanced the protein stability of TFR1 in MPC5 cells. ( A) , Western blot was performed to assess the protein levels of SIRT5, GPX4, SLC7A11, and TFR1 after SIRT5 inhibition in MPC5 cells ( N = 3); ( B) , IP and Western blot were performed to detect the protein levels of SIRT5, GPX4, SLC7A11, and TFR1 succinylation in MPC5 cells ( N = 3); ( C) , The protein levels of TFR1 and TFR1-suc in MPC5 cells after mutations at K242, K384 and K626 sites of TFR1 were analyzed by Western blot ( N = 3); ( D) , Co-IP and ( E) , PLA assay were used to analyze the interaction between SIRT5 and TFR1 in MPC5 cells ( N = 3); ( F) , The MPC5 cells in each group were treated with CHX, then the protein level of SIRT5 and TFR1 in each group was assayed by Western blot at different time points (0, 6, 12, 18, and 24 h) ( N = 3). ** p < 0.01.

Article Snippet: MPC5 cells were transfected with SIRT5-targeting short hairpin (sh) RNA (shSIRT5), shRNA negative control (shNC), pcDNA3.1 empty vector, or transferrin receptor 1 (TFR1) overexpression plasmid (pcDNA3.1-TFR1) using Lipofectamine 3000 (#L3000001; Thermo Fisher Scientific, Waltham, MA, USA) following the manufacturer’s protocol.

Techniques: Western Blot, Inhibition, Co-Immunoprecipitation Assay

Journal: Scientific Reports

Article Title: Quercetin-4’-O-β-D-glucopyranoside inhibits ferroptosis though SIRT5-mediated desuccinylation of TFR1 in diabetic nephropathy

doi: 10.1038/s41598-026-41148-4

Figure Lengend Snippet: Overexpression of TFR1 reversed the inhibited ferroptosis induced by SIRT5 overexpression in MPC5 cells. ( A) , RT-qPCR was performed to assess the mRNA level of SIRT5 after SIRT5 overexpression in MPC5 cells ( N = 3); ( B) , The mRNA and protein levels of TFR1 after transfection of TFR1 overexpression vectors into MPC5 cells were analyzed by RT-qPCR and Western blot ( N = 3); ( C) , CCK-8 was performed to analyze the cell ability of each group ( N = 3); ( D) , Flow cytometry was performed to analyze the relative fluorescence intensity of lipid peroxidation in each group ( N = 3); Cellular ( E) , SOD, ( F) , CAT, ( G) , MDA, and ( H) , iron contents in each group were determined by commercial kits ( N = 3); ( I) , Western blot was performed to assess the protein levels of SIRT5, GPX4, SLC7A11, and TFR1 in each group ( N = 3). ** p < 0.01.

Article Snippet: MPC5 cells were transfected with SIRT5-targeting short hairpin (sh) RNA (shSIRT5), shRNA negative control (shNC), pcDNA3.1 empty vector, or transferrin receptor 1 (TFR1) overexpression plasmid (pcDNA3.1-TFR1) using Lipofectamine 3000 (#L3000001; Thermo Fisher Scientific, Waltham, MA, USA) following the manufacturer’s protocol.

Techniques: Over Expression, Quantitative RT-PCR, Transfection, Western Blot, CCK-8 Assay, Flow Cytometry, Fluorescence

Journal: Scientific Reports

Article Title: Quercetin-4’-O-β-D-glucopyranoside inhibits ferroptosis though SIRT5-mediated desuccinylation of TFR1 in diabetic nephropathy

doi: 10.1038/s41598-026-41148-4

Figure Lengend Snippet: QODG reversed the renal injury induced by DN. ( A) , Representative images of H&E staining of the each group kidney sections (scale bar: 20 μm) ( N = 6); ( B) , Scr, ( C) , BUN, and ( D) , UAER values of the each group mice were shown ( N = 6); Commercial kits were used to assess the serum ( E) , ROS, ( F) , MDA, ( G) , CAT, ( H) , SOD, ( I) , iron, and ( J) , GSH contents in each group ( N = 6); ( K) , Western blot was performed to assess the protein levels of GPX4, SLC7A11, and TFR1 in the kidney tissues of each group mice ( N = 6); ( L) , IHC was performed to assess the expression levels of GPX4, SLC7A11, and TFR1 in the kidney tissues of each group mice (scale bar: 20 μm) ( N = 6). ** p < 0.01.

Article Snippet: MPC5 cells were transfected with SIRT5-targeting short hairpin (sh) RNA (shSIRT5), shRNA negative control (shNC), pcDNA3.1 empty vector, or transferrin receptor 1 (TFR1) overexpression plasmid (pcDNA3.1-TFR1) using Lipofectamine 3000 (#L3000001; Thermo Fisher Scientific, Waltham, MA, USA) following the manufacturer’s protocol.

Techniques: Staining, Western Blot, Expressing