Journal: Scientific Reports

Article Title: Rubidium ions as a novel therapeutic approach for glioblastoma

doi: 10.1038/s41598-025-97688-8

Figure Lengend Snippet: Rb⁺ significantly inhibited cell proliferation and migration in U87-MG and U251 cells. ( A ) CCK8 assay showed that Rb⁺ inhibits proliferation of U251 and U87-MG cells. ( B ) Transmission electron microscope combined with energy dispersive spectrometry confirmed that Rb⁺ could penetrate and accumulate within human glioblastoma cells. ( C ) Wound healing assays confirmed that Rb⁺ significantly inhibited glioblastoma cell migration. The data are presented as mean ± SD, ** P < 0.01, *** P < 0.001, **** P < 0.0001 vs. control group.

Article Snippet: The human GBM cell lines U87 and U251 (Wuhan Pricella Biotechnology) were cultured in DMEM (Gibco, CA, USA) supplemented with 10% FBS (PWL001-3, Meiluncell, China), 100 U/mL penicillin, and 100 µg/mL streptomycin.

Techniques: Migration, CCK-8 Assay, Transmission Assay, Microscopy, Control

Journal: Scientific Reports

Article Title: Rubidium ions as a novel therapeutic approach for glioblastoma

doi: 10.1038/s41598-025-97688-8

Figure Lengend Snippet: Rb⁺ significantly inhibited cell invasion and colony formation in U87-MG and U251cells. ( A ) Colony formation assays revealed that Rb⁺ significantly inhibited the colony formation of U251 and U87-MG cells. ( B ) Morphological changes in cells treated with Rb⁺. ( C ) Transwell assays confirmed that Rb⁺ significantly inhibited glioblastoma cell invasion. The data are presented as mean ± SD, ** P < 0.01, *** P < 0.001, **** P < 0.0001 vs. control group.

Article Snippet: The human GBM cell lines U87 and U251 (Wuhan Pricella Biotechnology) were cultured in DMEM (Gibco, CA, USA) supplemented with 10% FBS (PWL001-3, Meiluncell, China), 100 U/mL penicillin, and 100 µg/mL streptomycin.

Techniques: Control

Journal: Scientific Reports

Article Title: Rubidium ions as a novel therapeutic approach for glioblastoma

doi: 10.1038/s41598-025-97688-8

Figure Lengend Snippet: Rb⁺ induced apoptosis of glioblastoma cell and cause G2/M cell cycle arrest in vitro. ( A ) Transmission electron microscope revealed typical apoptotic characteristics in U87 and U251 cells following Rb⁺ treatment. ( B ) Flow cytometry showed a significant increase in DNA fragmentation in both cell lines, which was concentration-dependent. ( C ) Annexin V/PI staining confirmed that Rb⁺ effectively induced apoptosis in a dose-dependent manner. ( D ) Rb⁺ caused dose-dependent G2/M phase arrest in both U87 and U251 cells. ( E ) Western blot analysis showed that Rb⁺ upregulated BAX and Caspase-3 while downregulating BCL-2, confirming their role in promoting apoptosis. The original blots are presented in Supplementary Fig.S3. The samples derive from the same experiment and that blots were processed in parallel.

Article Snippet: The human GBM cell lines U87 and U251 (Wuhan Pricella Biotechnology) were cultured in DMEM (Gibco, CA, USA) supplemented with 10% FBS (PWL001-3, Meiluncell, China), 100 U/mL penicillin, and 100 µg/mL streptomycin.

Techniques: In Vitro, Transmission Assay, Microscopy, Flow Cytometry, Concentration Assay, Staining, Western Blot

Journal: Scientific Reports

Article Title: Rubidium ions as a novel therapeutic approach for glioblastoma

doi: 10.1038/s41598-025-97688-8

Figure Lengend Snippet: Effect of Rb⁺ on the PI3K-AKT-mTOR pathway: RNA sequencing and Western blot verification. ( A ) A volcano map showed a large number of up-regulated and down-regulated genes in rubidium chlorine treated U87 and U251 cells compared to controls. ( B, C ) Heat maps ( B ) and plot of principal component analysis ( C ) further showed differentially expressed genes between the treatment and control groups. ( D ) KEGG pathway enrichment analysis revealed that Rb⁺ treatment significantly affected several apoptotic and cell cycle pathways. ( E ) Western blot analysis confirmed that Rb⁺ promoted apoptosis by downregulating the PI3K/AKT/mTOR signaling pathway and its downstream targets. The original blots are presented in Supplementary Fig.S4. The samples derive from the same experiment and that blots were processed in parallel.

Article Snippet: The human GBM cell lines U87 and U251 (Wuhan Pricella Biotechnology) were cultured in DMEM (Gibco, CA, USA) supplemented with 10% FBS (PWL001-3, Meiluncell, China), 100 U/mL penicillin, and 100 µg/mL streptomycin.

Techniques: RNA Sequencing, Western Blot, Control

Journal: International Journal of Molecular Sciences

Article Title: FDX1 Regulates the Phosphorylation of ATM, DNA-PKcs Akt, and EGFR and Affects Radioresistance Under Severe Hypoxia in the Glioblastoma Cell Line T98G

doi: 10.3390/ijms26073378

Figure Lengend Snippet: Induction of FDX1 expression under severe hypoxia and the effects of FDX1 knockdown on the activation of Akt and EGFR. ( A ) The effects of severe hypoxia on the expression and/or phosphorylation of FDX1, Akt, and EGFR were investigated in T98G cells with FDX1 knockdown. After 48 h of siRNA treatment (siCtrl or siFDX1), cells were cultured for 18 h under either severe hypoxia or normoxia and then subjected to Western blot analysis using the indicated antibodies. β-actin served as an internal control. ( B ) Quantification of the effect of siFDX1 on FDX1 expression levels under severe hypoxia (with siCtrl set to 1). ( C ) Quantification of the effect of siFDX1 on the phosphorylation levels of Akt and EGFR under severe hypoxia (with siCtrl set to 1). Data are expressed as mean ± SD from three independent experiments ( n = 3); ns: not significant, * p < 0.05, ** p < 0.01, *** p < 0.001 (Student’s t -test). “Nx” represents normoxia, and “Hx” represents severe hypoxia.

Article Snippet: Human glioblastoma (GBM) cell lines T98G and A172 were obtained from the American Type Culture Collection (Rockville, MD, USA).

Techniques: Expressing, Knockdown, Activation Assay, Cell Culture, Western Blot, Control

Journal: International Journal of Molecular Sciences

Article Title: FDX1 Regulates the Phosphorylation of ATM, DNA-PKcs Akt, and EGFR and Affects Radioresistance Under Severe Hypoxia in the Glioblastoma Cell Line T98G

doi: 10.3390/ijms26073378

Figure Lengend Snippet: Effects of FDX1 knockdown on radioresistance and cell cycle distribution under severe hypoxia. Radiosensitivity under severe hypoxia was assessed in T98G cells with FDX1 knockdown. After 48 h of siRNA treatment (siCtrl or siFDX1), cells were cultured for 18 h under either severe hypoxia or normoxia. Subsequently, cells were irradiated with 0 or 6 Gy X-rays while maintaining hypoxia. After irradiation, cells were cultured for an additional 4 h under the same conditions before being used for the colony formation assay. ( A ) Box plots showing the colony formation assay results (6 Gy). Values are presented as mean ± SD ( n = 3). ns: not significant, ** p < 0.01 (Student’s t -test). Results are also shown in . ( B ) The effect of FDX1 knockdown on cell cycle distribution in T98G cells under severe hypoxia was analyzed by flow cytometry. Values are presented as mean ± SD from three independent experiments ( n = 3); ns: not significant (Dunnett’s test). “Nx” represents normoxia, and “Hx” represents severe hypoxia.

Article Snippet: Human glioblastoma (GBM) cell lines T98G and A172 were obtained from the American Type Culture Collection (Rockville, MD, USA).

Techniques: Knockdown, Cell Culture, Irradiation, Colony Assay, Flow Cytometry

Journal: International Journal of Molecular Sciences

Article Title: FDX1 Regulates the Phosphorylation of ATM, DNA-PKcs Akt, and EGFR and Affects Radioresistance Under Severe Hypoxia in the Glioblastoma Cell Line T98G

doi: 10.3390/ijms26073378

Figure Lengend Snippet: Effects of FDX1 knockdown on the expression and activation of DNA double-strand break repair enzymes under severe hypoxia. The effects of severe hypoxia on the expression and phosphorylation of DNA-PKcs and ATM were examined in T98G cells with FDX1 knockdown. After 48 h of siRNA treatment (siCtrl or siFDX1), cells were cultured for 18 h under either severe hypoxia or normoxia. ( A ) Cells were then processed for Western blot analysis using the indicated antibodies. β-actin was used as an internal control. ( B ) Quantification of the effect of siFDX1 on the phosphorylation levels of DNA-PKcs and ATM under severe hypoxia (with siCtrl set to 1). Data are expressed as mean ± SD from three independent experiments. ** p < 0.01 (Student’s t -test). “Nx” represents normoxia, and “Hx” represents severe hypoxia. ( C ) Representative confocal microscopy images showing the effect of FDX1 knockdown on the subcellular localization of phosphorylated DNA-PKcs under severe hypoxia.

Article Snippet: Human glioblastoma (GBM) cell lines T98G and A172 were obtained from the American Type Culture Collection (Rockville, MD, USA).

Techniques: Knockdown, Expressing, Activation Assay, Cell Culture, Western Blot, Control, Confocal Microscopy

Journal: International Journal of Molecular Sciences

Article Title: FDX1 Regulates the Phosphorylation of ATM, DNA-PKcs Akt, and EGFR and Affects Radioresistance Under Severe Hypoxia in the Glioblastoma Cell Line T98G

doi: 10.3390/ijms26073378

Figure Lengend Snippet: Effects of FDX1 knockdown on ATM, DNA-PKcs, and Akt expression and activation following X-ray irradiation under severe hypoxia. The impact of X-ray irradiation under severe hypoxia on the expression and phosphorylation of Akt, DNA-PKcs, and ATM was examined in T98G cells with FDX1 knockdown. After 48 h of siRNA treatment (siCtrl or siFDX1), the cells were cultured for 18 h under either severe hypoxia or normoxia. Following irradiation with 6 Gy of X-rays, the cells were cultured for 4 h and then subjected to Western blot analysis with antibodies indicated. β-actin as an internal control. “Nx” represents normoxia, while “Hx” represents severe hypoxia.

Article Snippet: Human glioblastoma (GBM) cell lines T98G and A172 were obtained from the American Type Culture Collection (Rockville, MD, USA).

Techniques: Knockdown, Expressing, Activation Assay, Irradiation, Cell Culture, Western Blot, Control

Journal: International Journal of Molecular Sciences

Article Title: FDX1 Regulates the Phosphorylation of ATM, DNA-PKcs Akt, and EGFR and Affects Radioresistance Under Severe Hypoxia in the Glioblastoma Cell Line T98G

doi: 10.3390/ijms26073378

Figure Lengend Snippet: Effects of FDX1, HIF-1α, and AMPKα knockdown on the expression of each under severe hypoxia. ( A ) The effects of severe hypoxia on HIF-1α protein levels and AMPKα expression were examined using T98G cells with FDX1 knockdown. ( B ) The effect of HIF-1α knockdown on FDX1 expression. ( C ) The effect of AMPKα knockdown on FDX1 expression. β-actin was used as an internal control. “Nx” represents normoxia, and “Hx” represents severe hypoxia.

Article Snippet: Human glioblastoma (GBM) cell lines T98G and A172 were obtained from the American Type Culture Collection (Rockville, MD, USA).

Techniques: Knockdown, Expressing, Control