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(A) Glioma cell survival at 24 h after SS treatment. (B) Glioma cell survival after pretreatment with DFO, FAC, Fer‐1, and Lip‐1, respectively, for 1 h before SS treatment. (C–F) Intracellular ROS, MDA, GSH/GSSG, and ferrous ion levels at 6, 12, and 24 h after SS treatment, with and without DFO, Fer‐1, Lip‐1, and FAC pretreatment for 1 h before SS treatment, respectively. (G) Glioma cell survival after SS treatment, with and without pretreatment with NAC and GSH for 1 h before SS treatment, respectively. (H) Western blots of ferroptosis‐related protein expression after SS treatment. Cell survival was assessed by the <t>CCK‐8</t> assay. (I) Western blots of ATF3 expression in the cytoplasm and nuclei of SS‐treated cells. (J) Viability of cells treated with siATF3 followed by SS. (K–N) Ferrous ion, MDA, GSH/GSSG, and ROS levels in cells treated with siATF3 followed by SS. (O) Confocal microscopic images of ATF3 immunofluorescence in U251 cells treated with SS. (P, Q) Confocal microscopy images and fluorescence intensity measurements of mitochondrial superoxide in SS‐ and siATF3‐treated U251 and T98 cells. * p < 0.01 versus control, # p < 0.01 versus treatment group.
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(A) Glioma cell survival at 24 h after SS treatment. (B) Glioma cell survival after pretreatment with DFO, FAC, Fer‐1, and Lip‐1, respectively, for 1 h before SS treatment. (C–F) Intracellular ROS, MDA, GSH/GSSG, and ferrous ion levels at 6, 12, and 24 h after SS treatment, with and without DFO, Fer‐1, Lip‐1, and FAC pretreatment for 1 h before SS treatment, respectively. (G) Glioma cell survival after SS treatment, with and without pretreatment with NAC and GSH for 1 h before SS treatment, respectively. (H) Western blots of ferroptosis‐related protein expression after SS treatment. Cell survival was assessed by the <t>CCK‐8</t> assay. (I) Western blots of ATF3 expression in the cytoplasm and nuclei of SS‐treated cells. (J) Viability of cells treated with siATF3 followed by SS. (K–N) Ferrous ion, MDA, GSH/GSSG, and ROS levels in cells treated with siATF3 followed by SS. (O) Confocal microscopic images of ATF3 immunofluorescence in U251 cells treated with SS. (P, Q) Confocal microscopy images and fluorescence intensity measurements of mitochondrial superoxide in SS‐ and siATF3‐treated U251 and T98 cells. * p < 0.01 versus control, # p < 0.01 versus treatment group.
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(A) Glioma cell survival at 24 h after SS treatment. (B) Glioma cell survival after pretreatment with DFO, FAC, Fer‐1, and Lip‐1, respectively, for 1 h before SS treatment. (C–F) Intracellular ROS, MDA, GSH/GSSG, and ferrous ion levels at 6, 12, and 24 h after SS treatment, with and without DFO, Fer‐1, Lip‐1, and FAC pretreatment for 1 h before SS treatment, respectively. (G) Glioma cell survival after SS treatment, with and without pretreatment with NAC and GSH for 1 h before SS treatment, respectively. (H) Western blots of ferroptosis‐related protein expression after SS treatment. Cell survival was assessed by the <t>CCK‐8</t> assay. (I) Western blots of ATF3 expression in the cytoplasm and nuclei of SS‐treated cells. (J) Viability of cells treated with siATF3 followed by SS. (K–N) Ferrous ion, MDA, GSH/GSSG, and ROS levels in cells treated with siATF3 followed by SS. (O) Confocal microscopic images of ATF3 immunofluorescence in U251 cells treated with SS. (P, Q) Confocal microscopy images and fluorescence intensity measurements of mitochondrial superoxide in SS‐ and siATF3‐treated U251 and T98 cells. * p < 0.01 versus control, # p < 0.01 versus treatment group.
Cell Counting Kit 8, supplied by Beijing Solarbio Science, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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cell counting kit 8 cck 8  (Beijing Solarbio Science)
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(A) Glioma cell survival at 24 h after SS treatment. (B) Glioma cell survival after pretreatment with DFO, FAC, Fer‐1, and Lip‐1, respectively, for 1 h before SS treatment. (C–F) Intracellular ROS, MDA, GSH/GSSG, and ferrous ion levels at 6, 12, and 24 h after SS treatment, with and without DFO, Fer‐1, Lip‐1, and FAC pretreatment for 1 h before SS treatment, respectively. (G) Glioma cell survival after SS treatment, with and without pretreatment with NAC and GSH for 1 h before SS treatment, respectively. (H) Western blots of ferroptosis‐related protein expression after SS treatment. Cell survival was assessed by the <t>CCK‐8</t> assay. (I) Western blots of ATF3 expression in the cytoplasm and nuclei of SS‐treated cells. (J) Viability of cells treated with siATF3 followed by SS. (K–N) Ferrous ion, MDA, GSH/GSSG, and ROS levels in cells treated with siATF3 followed by SS. (O) Confocal microscopic images of ATF3 immunofluorescence in U251 cells treated with SS. (P, Q) Confocal microscopy images and fluorescence intensity measurements of mitochondrial superoxide in SS‐ and siATF3‐treated U251 and T98 cells. * p < 0.01 versus control, # p < 0.01 versus treatment group.
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cck 8  (Beijing Solarbio Science)
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(A) Glioma cell survival at 24 h after SS treatment. (B) Glioma cell survival after pretreatment with DFO, FAC, Fer‐1, and Lip‐1, respectively, for 1 h before SS treatment. (C–F) Intracellular ROS, MDA, GSH/GSSG, and ferrous ion levels at 6, 12, and 24 h after SS treatment, with and without DFO, Fer‐1, Lip‐1, and FAC pretreatment for 1 h before SS treatment, respectively. (G) Glioma cell survival after SS treatment, with and without pretreatment with NAC and GSH for 1 h before SS treatment, respectively. (H) Western blots of ferroptosis‐related protein expression after SS treatment. Cell survival was assessed by the <t>CCK‐8</t> assay. (I) Western blots of ATF3 expression in the cytoplasm and nuclei of SS‐treated cells. (J) Viability of cells treated with siATF3 followed by SS. (K–N) Ferrous ion, MDA, GSH/GSSG, and ROS levels in cells treated with siATF3 followed by SS. (O) Confocal microscopic images of ATF3 immunofluorescence in U251 cells treated with SS. (P, Q) Confocal microscopy images and fluorescence intensity measurements of mitochondrial superoxide in SS‐ and siATF3‐treated U251 and T98 cells. * p < 0.01 versus control, # p < 0.01 versus treatment group.
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cck 8 solution  (Beijing Solarbio Science)
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(A) Glioma cell survival at 24 h after SS treatment. (B) Glioma cell survival after pretreatment with DFO, FAC, Fer‐1, and Lip‐1, respectively, for 1 h before SS treatment. (C–F) Intracellular ROS, MDA, GSH/GSSG, and ferrous ion levels at 6, 12, and 24 h after SS treatment, with and without DFO, Fer‐1, Lip‐1, and FAC pretreatment for 1 h before SS treatment, respectively. (G) Glioma cell survival after SS treatment, with and without pretreatment with NAC and GSH for 1 h before SS treatment, respectively. (H) Western blots of ferroptosis‐related protein expression after SS treatment. Cell survival was assessed by the <t>CCK‐8</t> assay. (I) Western blots of ATF3 expression in the cytoplasm and nuclei of SS‐treated cells. (J) Viability of cells treated with siATF3 followed by SS. (K–N) Ferrous ion, MDA, GSH/GSSG, and ROS levels in cells treated with siATF3 followed by SS. (O) Confocal microscopic images of ATF3 immunofluorescence in U251 cells treated with SS. (P, Q) Confocal microscopy images and fluorescence intensity measurements of mitochondrial superoxide in SS‐ and siATF3‐treated U251 and T98 cells. * p < 0.01 versus control, # p < 0.01 versus treatment group.
Cck 8 Solution, supplied by Beijing Solarbio Science, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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(A) Glioma cell survival at 24 h after SS treatment. (B) Glioma cell survival after pretreatment with DFO, FAC, Fer‐1, and Lip‐1, respectively, for 1 h before SS treatment. (C–F) Intracellular ROS, MDA, GSH/GSSG, and ferrous ion levels at 6, 12, and 24 h after SS treatment, with and without DFO, Fer‐1, Lip‐1, and FAC pretreatment for 1 h before SS treatment, respectively. (G) Glioma cell survival after SS treatment, with and without pretreatment with NAC and GSH for 1 h before SS treatment, respectively. (H) Western blots of ferroptosis‐related protein expression after SS treatment. Cell survival was assessed by the <t>CCK‐8</t> assay. (I) Western blots of ATF3 expression in the cytoplasm and nuclei of SS‐treated cells. (J) Viability of cells treated with siATF3 followed by SS. (K–N) Ferrous ion, MDA, GSH/GSSG, and ROS levels in cells treated with siATF3 followed by SS. (O) Confocal microscopic images of ATF3 immunofluorescence in U251 cells treated with SS. (P, Q) Confocal microscopy images and fluorescence intensity measurements of mitochondrial superoxide in SS‐ and siATF3‐treated U251 and T98 cells. * p < 0.01 versus control, # p < 0.01 versus treatment group.
Cck 8 Kit, supplied by Beyotime, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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cell counting kit 8 cck 8  (Beyotime)
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(A) Glioma cell survival at 24 h after SS treatment. (B) Glioma cell survival after pretreatment with DFO, FAC, Fer‐1, and Lip‐1, respectively, for 1 h before SS treatment. (C–F) Intracellular ROS, MDA, GSH/GSSG, and ferrous ion levels at 6, 12, and 24 h after SS treatment, with and without DFO, Fer‐1, Lip‐1, and FAC pretreatment for 1 h before SS treatment, respectively. (G) Glioma cell survival after SS treatment, with and without pretreatment with NAC and GSH for 1 h before SS treatment, respectively. (H) Western blots of ferroptosis‐related protein expression after SS treatment. Cell survival was assessed by the <t>CCK‐8</t> assay. (I) Western blots of ATF3 expression in the cytoplasm and nuclei of SS‐treated cells. (J) Viability of cells treated with siATF3 followed by SS. (K–N) Ferrous ion, MDA, GSH/GSSG, and ROS levels in cells treated with siATF3 followed by SS. (O) Confocal microscopic images of ATF3 immunofluorescence in U251 cells treated with SS. (P, Q) Confocal microscopy images and fluorescence intensity measurements of mitochondrial superoxide in SS‐ and siATF3‐treated U251 and T98 cells. * p < 0.01 versus control, # p < 0.01 versus treatment group.
Cell Counting Kit 8 Cck 8, supplied by Beyotime, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Image Search Results


(A) Glioma cell survival at 24 h after SS treatment. (B) Glioma cell survival after pretreatment with DFO, FAC, Fer‐1, and Lip‐1, respectively, for 1 h before SS treatment. (C–F) Intracellular ROS, MDA, GSH/GSSG, and ferrous ion levels at 6, 12, and 24 h after SS treatment, with and without DFO, Fer‐1, Lip‐1, and FAC pretreatment for 1 h before SS treatment, respectively. (G) Glioma cell survival after SS treatment, with and without pretreatment with NAC and GSH for 1 h before SS treatment, respectively. (H) Western blots of ferroptosis‐related protein expression after SS treatment. Cell survival was assessed by the CCK‐8 assay. (I) Western blots of ATF3 expression in the cytoplasm and nuclei of SS‐treated cells. (J) Viability of cells treated with siATF3 followed by SS. (K–N) Ferrous ion, MDA, GSH/GSSG, and ROS levels in cells treated with siATF3 followed by SS. (O) Confocal microscopic images of ATF3 immunofluorescence in U251 cells treated with SS. (P, Q) Confocal microscopy images and fluorescence intensity measurements of mitochondrial superoxide in SS‐ and siATF3‐treated U251 and T98 cells. * p < 0.01 versus control, # p < 0.01 versus treatment group.

Journal: CNS Neuroscience & Therapeutics

Article Title: ATF3 Promotes Solasonine‐Triggered Ferroptosis in Glioma Cells by Suppressing Glycolysis

doi: 10.1002/cns.71226

Figure Lengend Snippet: (A) Glioma cell survival at 24 h after SS treatment. (B) Glioma cell survival after pretreatment with DFO, FAC, Fer‐1, and Lip‐1, respectively, for 1 h before SS treatment. (C–F) Intracellular ROS, MDA, GSH/GSSG, and ferrous ion levels at 6, 12, and 24 h after SS treatment, with and without DFO, Fer‐1, Lip‐1, and FAC pretreatment for 1 h before SS treatment, respectively. (G) Glioma cell survival after SS treatment, with and without pretreatment with NAC and GSH for 1 h before SS treatment, respectively. (H) Western blots of ferroptosis‐related protein expression after SS treatment. Cell survival was assessed by the CCK‐8 assay. (I) Western blots of ATF3 expression in the cytoplasm and nuclei of SS‐treated cells. (J) Viability of cells treated with siATF3 followed by SS. (K–N) Ferrous ion, MDA, GSH/GSSG, and ROS levels in cells treated with siATF3 followed by SS. (O) Confocal microscopic images of ATF3 immunofluorescence in U251 cells treated with SS. (P, Q) Confocal microscopy images and fluorescence intensity measurements of mitochondrial superoxide in SS‐ and siATF3‐treated U251 and T98 cells. * p < 0.01 versus control, # p < 0.01 versus treatment group.

Article Snippet: SS and cell counting kit‐8 (CCK‐8) kits were purchased from TargetMol (China).

Techniques: Western Blot, Expressing, CCK-8 Assay, Confocal, Immunofluorescence, Microscopy, Fluorescence, Control