ag1  (MedChemExpress)

Journal: Cell Death & Disease

Article Title: Post-irradiation dietary restriction impairs hematopoiesis via inhibition of the pentose phosphate pathway in hematopoietic stem and progenitor cells

doi: 10.1038/s41419-025-08249-w

Figure Lengend Snippet: A – E Mice were subjected to 4.5 Gy X-ray irradiation and subsequently fed with AL diet or DR diet. BM cells or c-Kit + HSPCs were harvested at the indicated time points for q RT-PCR analysis. NIR mice receiving an AL diet were monitored as a control. Relative expression of indicated genes involving DNA damage and repair were analyzed, with β-actin as the internal control ( n = 5 mice per group from 1 experiment representative of 2 independent experiments). F , G Apoptosis rates in HSCs at indicated time points post-irradiation and in NIR mice, as determined by FACS ( F ). Representative FACS plots at day 1 post-irradiation ( G ) ( n = 5 mice per group from 1 experiment, representative of 2 independent experiments). H , I Quantification and representative images of γH2AX and 53BP1 colocalization in c-Kit + HSPCs at indicated time points post-irradiation and before irradiation, determined by immunofluorescent staining (scale bar: 10 μm) ( n = 5 mice per group from 1 experiment representative of 2 independent experiments). J – L Quantification and representative images of comet assay tail DNA in c-Kit + HSPCs at indicated time points post-irradiation and before irradiation (scale bar: 10 μm) ( n = 5 mice per group from 1 experiment, representative of 2 independent experiments). M Relative expression of G6PD mRNA in BM cells of AL and DR mice at the indicated time points post-irradiation and NIR AL mice, analyzed by qRT-PCR with β-actin as the internal control ( n = 5 mice per group from 1 experiment representative of 2 independent experiments). Dynamic measurements by ELISA of G6PD enzyme activity ( N ) and NADPH ( O ) content in BM cells of AL and DR mice at the indicated time points under IR or no-conditions conditions. P G6PD enzyme activity and Q NADPH content in BM cells of AL and DR mice given 5% sucrose water or normal drinking water at NIR/IR 14 days ( n = 5 mice per group from 1 experiment representative of 2 independent experiments). R Average fluorescence intensity of ROS in BM cells of AL and DR mice at indicated time points under non-irradiated or irradiated conditions ( n = 5 mice per group from 1 experiment representative of 2 independent experiments). Relative expression of indicated genes involving ROS in BM cells ( S ) and c-Kit + HSPCs ( T ) of AL and DR mice at indicated time points post-irradiation, analyzed by qRT-PCR with NIR AL mice as a control ( n = 5 mice per group from 1 experiment representative of 2 independent experiments). U and V Quantification and representative images of Nrf2 foci in the nucleus of BM cells of AL and DR mice at 5 h post-irradiation and under non-irradiated conditions, determined by immunofluorescent staining (scale bar: 10 μm) ( n = 5 mice per group from 1 experiment representative of 2 independent experiments). Results were displayed as mean ± SD;. ns, not significant; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001 by one-way ANOVA test and Two-way ANOVA test with Tukey’s multiple comparisons test. HSPCs: hematopoietic stem and progenitor cells. The purple symbols indicate statistical significance between the non-irradiated groups. The green symbols indicate statistical significance between the irradiated groups.

Article Snippet: The G6PD activator AG1 was purchased from MedChemExpress Co., Ltd., China (CAS No. 421581-52-4; Lot number: HY-123962; purity: 99.54%), and was diluted in dimethyl sulfoxide (DMSO) for injection intraperitoneally every other day for 9 times post-irradiation.

Techniques: Irradiation, Reverse Transcription Polymerase Chain Reaction, Control, Expressing, Staining, Single Cell Gel Electrophoresis, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Activity Assay, Fluorescence

Journal: Cell Death & Disease

Article Title: Post-irradiation dietary restriction impairs hematopoiesis via inhibition of the pentose phosphate pathway in hematopoietic stem and progenitor cells

doi: 10.1038/s41419-025-08249-w

Figure Lengend Snippet: A Experimental scheme. Wild-type C57BL/6 mice (2 months old) were irradiated with 4.5 Gy X-rays and fed with an AL diet. Mice were then injected intraperitoneally with 10 mg/kg of 6-aminonicotinamide (6-AN) or saline daily for 7 days post-irradiation, and were continued on an AL diet until 1 month post-irradiation ( n = 5 mice per group from 1 experiment representative of 2 independent experiments). G6PD enzyme activity ( B ) and NADPH content ( C ) in BM cells of AL mice 1 month post-irradiation, analyzed using ELISA. WBC ( D ), Neutrophil ( E ), and lymphocyte ( F ) counts in PB of AL mice 1 month post-irradiation. G , H Spleen and thymus weights of AL mice 1 month post-irradiation. I Total BM cell counts of AL mice 1 month post-irradiation. FACS analysis of frequencies of B220 + lymphocytes in PB ( J ), spleen ( K ), BM ( L ); and frequencies of HSCs ( M ), CMPs ( O ), and GMPs ( P ), and representative FACS Plots of HSCs ( N ) and CMPs/GMPs/MEPs ( Q ) of AL mice 1 month post-irradiation. R and S Quantification and representative images of γH2AX and 53BP1 colocalization in BM cells of AL mice 1 month post-irradiation, determined by immunofluorescent staining (scale bar: 10 μm). T–A’ Chimerism of donor-derived cells in the PB, including WBCs ( T ), B cells ( V ), T cells ( X ), and myeloid cells ( Z ) chimerism after transplantation at indicated time points and representative FACS plot of WBCs ( U ), B cells ( W ), T cells ( Y ), myeloid cells ( A’ ) at 4 months post-transplantation. Results were displayed as mean ± SD;. ns, not significant; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001 by unpaired two-tailed Student’s t -test and Two-way ANOVA test with Tukey’s multiple comparisons test. IR AL + 6-AN: irradiated ad libitum group injected with 6-AN; IR AL + NS: irradiated ad libitum group injected with saline.

Article Snippet: The G6PD activator AG1 was purchased from MedChemExpress Co., Ltd., China (CAS No. 421581-52-4; Lot number: HY-123962; purity: 99.54%), and was diluted in dimethyl sulfoxide (DMSO) for injection intraperitoneally every other day for 9 times post-irradiation.

Techniques: Irradiation, Injection, Saline, Activity Assay, Enzyme-linked Immunosorbent Assay, Staining, Derivative Assay, Transplantation Assay, Two Tailed Test

Journal: Cell Death & Disease

Article Title: Post-irradiation dietary restriction impairs hematopoiesis via inhibition of the pentose phosphate pathway in hematopoietic stem and progenitor cells

doi: 10.1038/s41419-025-08249-w

Figure Lengend Snippet: A Experimental scheme. Wild-type C57BL/6 mice (2 months old) were irradiated with 4.5 Gy X-rays, fed with a DR diet. Mice were then injected intraperitoneally with 20 mg/kg of AG1 or DMSO every other day for 9 times post-irradiation, and were continued on DR until 1 month post-irradiation ( n = 5 mice per group from 1 experiment representative of 2 independent experiments). G6PD enzyme activity ( B ) and NADPH content ( C ) in BM cells of DR mice 1 month post-irradiation, analyzed using ELISA. WBC ( D ), Neutrophil ( E ), and lymphocyte ( F ) counts in PB of DR mice 1 month post-irradiation. G , H Spleen and thymus weights of DR mice 1 month post-irradiation. I Total BM cell counts of DR mice 1 month post-irradiation. FACS analysis of frequencies of B220 + lymphocytes in PB ( J ), spleen ( K ), BM ( L ); and frequencies of HSCs ( M ), CMPs ( O ) and GMPs ( P ), and representative FACS Plots of HSCs ( N ) and CMPs/GMPs/MEPs ( Q ) of DR mice 1 month post-irradiation. R and S Quantification and representative images of γH2AX and 53BP1 colocalization in BM cells of DR mice 1 month post-irradiation, determined by immunofluorescent staining (scale bar: 10 μm). Chimerism of donor-derived cells in the PB, including WBCs ( T ), B cells ( V ), T cells ( X ), and myeloid cells ( Z ) chimerism after transplantation at indicated time points and representative FACS plot of WBCs ( U ), B cells ( W ), T cells ( Y ), myeloid cells ( A’ ) at 4 month post-transplantation. Results were displayed as mean ± SD;. ns, not significant; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001 by unpaired two-tailed Student’s t test, and Two-way ANOVA test with Tukey’s multiple comparisons test. IR DR + AG1: irradiated dietary restriction group injected with AG1; IR DR + DMSO: irradiated dietary restriction group injected with DMSO.

Article Snippet: The G6PD activator AG1 was purchased from MedChemExpress Co., Ltd., China (CAS No. 421581-52-4; Lot number: HY-123962; purity: 99.54%), and was diluted in dimethyl sulfoxide (DMSO) for injection intraperitoneally every other day for 9 times post-irradiation.

Techniques: Irradiation, Injection, Activity Assay, Enzyme-linked Immunosorbent Assay, Staining, Derivative Assay, Transplantation Assay, Two Tailed Test

Journal: Cell Death & Disease

Article Title: Post-irradiation dietary restriction impairs hematopoiesis via inhibition of the pentose phosphate pathway in hematopoietic stem and progenitor cells

doi: 10.1038/s41419-025-08249-w

Figure Lengend Snippet: A Experimental scheme. Wild-type C57BL/6 mice (2 months old) were irradiated with 4.5 Gy X-rays, fed with either AL diet or DR diet or DR diet 1 week, then AL feeding 3 weeks (daily food intake restricted to 70% of the intake of age- and sex-matched AL mice). NIR mice receiving an AL or DR diet were also monitored as a control ( n = 5 mice per group from 1 experiment representative of 2 independent experiments. B and C G6PD enzyme activity ( B ) and NADPH content ( C ) in BM cells of RF mice 1 month post-irradiation, analyzed using ELISA. WBC ( D ), Neutrophil ( E ), and lymphocyte ( F ), RBC ( G ), platelet ( H ) counts in PB of RF mice 1 month post-irradiation. I , J Spleen and thymus weights of RF mice 1 month post-irradiation. K Total BM cell counts of RF mice 1 month post-irradiation. FACS analysis of frequencies of B220 + lymphocytes in PB ( L ), spleen ( M ), BM ( N ); and frequencies of HSCs ( O ), CMPs ( P ) and GMPs ( Q ), MEPs ( R ), and RMBMs ( S ) of RF mice 1 month post-irradiation. T Apoptosis rates in HSCs of RF mice 1 month post-irradiation. U Average fluorescence intensity of ROS in BM cells of RF mice 1 month post-irradiation. V and W Quantification and representative images of γ-H2AX and 53BP1 colocalization in BM cells of RF mice 1 month post-irradiation, determined by immunofluorescent staining (scale bar: 10 μm). Results were displayed as mean ± SD; ns, not significant; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001 by one-way ANOVA test with Tukey’s multiple comparisons test. IR RF: irradiated refeeding group.

Article Snippet: The G6PD activator AG1 was purchased from MedChemExpress Co., Ltd., China (CAS No. 421581-52-4; Lot number: HY-123962; purity: 99.54%), and was diluted in dimethyl sulfoxide (DMSO) for injection intraperitoneally every other day for 9 times post-irradiation.

Techniques: Irradiation, Control, Activity Assay, Enzyme-linked Immunosorbent Assay, Fluorescence, Staining