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

Journal: Frontiers in Genetics

Article Title: CELF1 promotes aerobic glycolysis and an aggressive phenotype in ER-positive breast cancer via GLUT1 regulation

doi: 10.3389/fgene.2025.1687066

Figure Lengend Snippet: CELF1 promotes breast cancer cell aerobic glycolysis in vitro . (A) Enriched Gene Ontology (GO) terms of significantly differentially expressed genes between CELF1-knocked-out MCF7 cells and wild-type MCF7 cells. (B) GSEA shows the enriched hallmarks pathways between CELF1-knocked-out MCF7 cells and wild-type MCF7 cells. (C) GSEA shows the molecules that are significantly altered in the glycolytic pathway. (D) The volcano plot of differentially expressed genes between CELF1-knocked-out MCF7 cells and wild-type MCF7 cells. (E) Differential mRNA expressions of GLUT1, HK2, and G6PD in CELF1-knocked-out MCF7 cells and wild-type MCF7 cells.

Article Snippet: These membranes were probed with monoclonal antibodies targeting CELF1, GLUT1, cyclin D1, cyclin B1, c-Myc, HK, G6PD, and GAPDH (Proteintech, Wuhan, China), as well as Bcl-2 and BAX (Abbkine, United States).

Techniques: In Vitro

Journal: Frontiers in Genetics

Article Title: CELF1 promotes aerobic glycolysis and an aggressive phenotype in ER-positive breast cancer via GLUT1 regulation

doi: 10.3389/fgene.2025.1687066

Figure Lengend Snippet: CELF1 governs aerobic glycolysis by regulating GLUT1 levels. (A) Relative RNA expressions of GLUT1 HK and G6PD in CELF1-KO MCF7 cells (upper) and CELF1-overexpressed SKBR3 cells (lower) measured using real-time PCR. Data are expressed as means ± SEM. * P < 0.05.** P < 0.01. *** P < 0.001. (B) Immunofluorescence of GLUT1 in CELF1-KO MCF7 cells and CELF1-overexpressed SKBR3 cells. Scale bar: 50 μm. (C,D) Protein levels of CELF1, HK, c-Myc, G6PD, and GLUT1 in MCF7 cells after the transfection of control and CELF1 shRNA were determined using Western blotting. Cell lysates were collected with or without pretreated 20% fetal bovine serum (FBS) for 4 h. GAPDH served as an internal control. (E,F) . Protein levels of CELF1, HK, c-Myc, G6PD, and GLUT1 in SKBR3 cells after the transfection of the pcDNA3.1 and pcDNA3.1–CELF1 vectors were determined using Western blotting. Cell lysates were collected with or without pretreated 20% FBS for 4 h. GAPDH served as an internal control.

Article Snippet: These membranes were probed with monoclonal antibodies targeting CELF1, GLUT1, cyclin D1, cyclin B1, c-Myc, HK, G6PD, and GAPDH (Proteintech, Wuhan, China), as well as Bcl-2 and BAX (Abbkine, United States).

Techniques: Real-time Polymerase Chain Reaction, Immunofluorescence, Transfection, Control, shRNA, Western Blot

Journal: Frontiers in Genetics

Article Title: CELF1 promotes aerobic glycolysis and an aggressive phenotype in ER-positive breast cancer via GLUT1 regulation

doi: 10.3389/fgene.2025.1687066

Figure Lengend Snippet: Mechanism diagram illustrating the involvement of CELF1 in aerobic glycolysis in breast cancer. It is well known that many aggressive tumors develop dysregulated metabolism; the glycolytic pathway was closely correlated to the vitality of tumors. Our transcriptomic analysis results suggest that CELF1 alterations impact the glycolysis process, and GLUT1 is the main molecule among all the volatile metabolites. Therefore, we focused on the genes related to aerobic glycolysis. As shown in , the expression of GLUT1 is substantially decreased in the CELF1-knocked-out group, and so is the expression of key enzymes HK and G6PD in the aerobic glycolysis process . In addition, knockout of CELF1 affects the expression of cyclin D1 and c-Myc , suggesting that the occurrence, invasion, and metastasis processed are changed accordingly in the tumor tissues.

Article Snippet: These membranes were probed with monoclonal antibodies targeting CELF1, GLUT1, cyclin D1, cyclin B1, c-Myc, HK, G6PD, and GAPDH (Proteintech, Wuhan, China), as well as Bcl-2 and BAX (Abbkine, United States).

Techniques: Expressing, Knock-Out