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Phosphorylation of DNA repair proteins after Mirin treatment of fibroblasts. ( A ) Detailed representative images of γH2AX in SBLF24 treated with 2 Gy IR. Images were acquired by Zeiss, ApoTome.2 and Arivis software, interval of z-stacks was 0.24 μm. ( B ) Representative images of fibroblasts (SBLF24) treated with 30 µM Mirin and additionally with 2 Gy IR three hours afterwards (Mirin + IR). γH2AX and tumor protein P53 (p53) serine (Ser)20 were stained 21 h after irradiation. p53 Ser15 and ATM <t>Ser1981</t> were stained three hours after irradiation. Presented are the target proteins in green, nuclei in blue (DAPI) and merged images. Scale bar represents 10 μm. ( C ) Quantification of γH2AX, ATM Ser1981, p53 Ser15, and p53 Ser20 foci/nucleus. SBLF24 were treated with 30 µM Mirin and three hours afterwards additionally with a single dose of 2 Gy IR (Mirin + IR), only 30 µM Mirin (Mirin), or only 2 Gy IR (IR). Controls were completely untreated (Co). Statistically significant differences between IR and Mirin + IR were calculated with two-tailed Mann-Whitney test; ns: not significant. Each value represents mean ± SD ( n = 4; except p53 Ser20, Mirin, n = 3; ATM Ser1981, IR, n = 3)
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Phosphorylation of DNA repair proteins after Mirin treatment of fibroblasts. ( A ) Detailed representative images of γH2AX in SBLF24 treated with 2 Gy IR. Images were acquired by Zeiss, ApoTome.2 and Arivis software, interval of z-stacks was 0.24 μm. ( B ) Representative images of fibroblasts (SBLF24) treated with 30 µM Mirin and additionally with 2 Gy IR three hours afterwards (Mirin + IR). γH2AX and tumor protein P53 (p53) serine (Ser)20 were stained 21 h after irradiation. p53 Ser15 and ATM <t>Ser1981</t> were stained three hours after irradiation. Presented are the target proteins in green, nuclei in blue (DAPI) and merged images. Scale bar represents 10 μm. ( C ) Quantification of γH2AX, ATM Ser1981, p53 Ser15, and p53 Ser20 foci/nucleus. SBLF24 were treated with 30 µM Mirin and three hours afterwards additionally with a single dose of 2 Gy IR (Mirin + IR), only 30 µM Mirin (Mirin), or only 2 Gy IR (IR). Controls were completely untreated (Co). Statistically significant differences between IR and Mirin + IR were calculated with two-tailed Mann-Whitney test; ns: not significant. Each value represents mean ± SD ( n = 4; except p53 Ser20, Mirin, n = 3; ATM Ser1981, IR, n = 3)
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Phosphorylation of DNA repair proteins after Mirin treatment of fibroblasts. ( A ) Detailed representative images of γH2AX in SBLF24 treated with 2 Gy IR. Images were acquired by Zeiss, ApoTome.2 and Arivis software, interval of z-stacks was 0.24 μm. ( B ) Representative images of fibroblasts (SBLF24) treated with 30 µM Mirin and additionally with 2 Gy IR three hours afterwards (Mirin + IR). γH2AX and tumor protein P53 (p53) serine (Ser)20 were stained 21 h after irradiation. p53 Ser15 and ATM <t>Ser1981</t> were stained three hours after irradiation. Presented are the target proteins in green, nuclei in blue (DAPI) and merged images. Scale bar represents 10 μm. ( C ) Quantification of γH2AX, ATM Ser1981, p53 Ser15, and p53 Ser20 foci/nucleus. SBLF24 were treated with 30 µM Mirin and three hours afterwards additionally with a single dose of 2 Gy IR (Mirin + IR), only 30 µM Mirin (Mirin), or only 2 Gy IR (IR). Controls were completely untreated (Co). Statistically significant differences between IR and Mirin + IR were calculated with two-tailed Mann-Whitney test; ns: not significant. Each value represents mean ± SD ( n = 4; except p53 Ser20, Mirin, n = 3; ATM Ser1981, IR, n = 3)
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Phosphorylation of DNA repair proteins after Mirin treatment of fibroblasts. ( A ) Detailed representative images of γH2AX in SBLF24 treated with 2 Gy IR. Images were acquired by Zeiss, ApoTome.2 and Arivis software, interval of z-stacks was 0.24 μm. ( B ) Representative images of fibroblasts (SBLF24) treated with 30 µM Mirin and additionally with 2 Gy IR three hours afterwards (Mirin + IR). γH2AX and tumor protein P53 (p53) serine (Ser)20 were stained 21 h after irradiation. p53 Ser15 and ATM <t>Ser1981</t> were stained three hours after irradiation. Presented are the target proteins in green, nuclei in blue (DAPI) and merged images. Scale bar represents 10 μm. ( C ) Quantification of γH2AX, ATM Ser1981, p53 Ser15, and p53 Ser20 foci/nucleus. SBLF24 were treated with 30 µM Mirin and three hours afterwards additionally with a single dose of 2 Gy IR (Mirin + IR), only 30 µM Mirin (Mirin), or only 2 Gy IR (IR). Controls were completely untreated (Co). Statistically significant differences between IR and Mirin + IR were calculated with two-tailed Mann-Whitney test; ns: not significant. Each value represents mean ± SD ( n = 4; except p53 Ser20, Mirin, n = 3; ATM Ser1981, IR, n = 3)
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Phosphorylation of DNA repair proteins after Mirin treatment of fibroblasts. ( A ) Detailed representative images of γH2AX in SBLF24 treated with 2 Gy IR. Images were acquired by Zeiss, ApoTome.2 and Arivis software, interval of z-stacks was 0.24 μm. ( B ) Representative images of fibroblasts (SBLF24) treated with 30 µM Mirin and additionally with 2 Gy IR three hours afterwards (Mirin + IR). γH2AX and tumor protein P53 (p53) serine (Ser)20 were stained 21 h after irradiation. p53 Ser15 and ATM <t>Ser1981</t> were stained three hours after irradiation. Presented are the target proteins in green, nuclei in blue (DAPI) and merged images. Scale bar represents 10 μm. ( C ) Quantification of γH2AX, ATM Ser1981, p53 Ser15, and p53 Ser20 foci/nucleus. SBLF24 were treated with 30 µM Mirin and three hours afterwards additionally with a single dose of 2 Gy IR (Mirin + IR), only 30 µM Mirin (Mirin), or only 2 Gy IR (IR). Controls were completely untreated (Co). Statistically significant differences between IR and Mirin + IR were calculated with two-tailed Mann-Whitney test; ns: not significant. Each value represents mean ± SD ( n = 4; except p53 Ser20, Mirin, n = 3; ATM Ser1981, IR, n = 3)
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Phosphorylation of DNA repair proteins after Mirin treatment of fibroblasts. ( A ) Detailed representative images of γH2AX in SBLF24 treated with 2 Gy IR. Images were acquired by Zeiss, ApoTome.2 and Arivis software, interval of z-stacks was 0.24 μm. ( B ) Representative images of fibroblasts (SBLF24) treated with 30 µM Mirin and additionally with 2 Gy IR three hours afterwards (Mirin + IR). γH2AX and tumor protein P53 (p53) serine (Ser)20 were stained 21 h after irradiation. p53 Ser15 and ATM <t>Ser1981</t> were stained three hours after irradiation. Presented are the target proteins in green, nuclei in blue (DAPI) and merged images. Scale bar represents 10 μm. ( C ) Quantification of γH2AX, ATM Ser1981, p53 Ser15, and p53 Ser20 foci/nucleus. SBLF24 were treated with 30 µM Mirin and three hours afterwards additionally with a single dose of 2 Gy IR (Mirin + IR), only 30 µM Mirin (Mirin), or only 2 Gy IR (IR). Controls were completely untreated (Co). Statistically significant differences between IR and Mirin + IR were calculated with two-tailed Mann-Whitney test; ns: not significant. Each value represents mean ± SD ( n = 4; except p53 Ser20, Mirin, n = 3; ATM Ser1981, IR, n = 3)
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Image Search Results


Phosphorylation of DNA repair proteins after Mirin treatment of fibroblasts. ( A ) Detailed representative images of γH2AX in SBLF24 treated with 2 Gy IR. Images were acquired by Zeiss, ApoTome.2 and Arivis software, interval of z-stacks was 0.24 μm. ( B ) Representative images of fibroblasts (SBLF24) treated with 30 µM Mirin and additionally with 2 Gy IR three hours afterwards (Mirin + IR). γH2AX and tumor protein P53 (p53) serine (Ser)20 were stained 21 h after irradiation. p53 Ser15 and ATM Ser1981 were stained three hours after irradiation. Presented are the target proteins in green, nuclei in blue (DAPI) and merged images. Scale bar represents 10 μm. ( C ) Quantification of γH2AX, ATM Ser1981, p53 Ser15, and p53 Ser20 foci/nucleus. SBLF24 were treated with 30 µM Mirin and three hours afterwards additionally with a single dose of 2 Gy IR (Mirin + IR), only 30 µM Mirin (Mirin), or only 2 Gy IR (IR). Controls were completely untreated (Co). Statistically significant differences between IR and Mirin + IR were calculated with two-tailed Mann-Whitney test; ns: not significant. Each value represents mean ± SD ( n = 4; except p53 Ser20, Mirin, n = 3; ATM Ser1981, IR, n = 3)

Journal: Cancer Cell International

Article Title: The inhibition of the MRN complex by Mirin radiosensitizes particularly HPV-negative HNSCC cell lines

doi: 10.1186/s12935-026-04218-1

Figure Lengend Snippet: Phosphorylation of DNA repair proteins after Mirin treatment of fibroblasts. ( A ) Detailed representative images of γH2AX in SBLF24 treated with 2 Gy IR. Images were acquired by Zeiss, ApoTome.2 and Arivis software, interval of z-stacks was 0.24 μm. ( B ) Representative images of fibroblasts (SBLF24) treated with 30 µM Mirin and additionally with 2 Gy IR three hours afterwards (Mirin + IR). γH2AX and tumor protein P53 (p53) serine (Ser)20 were stained 21 h after irradiation. p53 Ser15 and ATM Ser1981 were stained three hours after irradiation. Presented are the target proteins in green, nuclei in blue (DAPI) and merged images. Scale bar represents 10 μm. ( C ) Quantification of γH2AX, ATM Ser1981, p53 Ser15, and p53 Ser20 foci/nucleus. SBLF24 were treated with 30 µM Mirin and three hours afterwards additionally with a single dose of 2 Gy IR (Mirin + IR), only 30 µM Mirin (Mirin), or only 2 Gy IR (IR). Controls were completely untreated (Co). Statistically significant differences between IR and Mirin + IR were calculated with two-tailed Mann-Whitney test; ns: not significant. Each value represents mean ± SD ( n = 4; except p53 Ser20, Mirin, n = 3; ATM Ser1981, IR, n = 3)

Article Snippet: ATM Ser1981 , Phospho-ATM (Ser1981) (10H11.E12) Mouse mAb; catalogue number: 4526; host: mouse; dilution: 1:300 (Cell Signaling Technology, Inc., Danvers, USA) , Donkey anti-Rabbit IgG (H + L) Highly Cross-Adsorbed Secondary Antibody, Alexa FluorTM 488; catalogue number: A-21,206; host: donkey; dilution: 1:200 (Invitrogen, Thermo Fisher Scientific Inc., Waltham, USA).

Techniques: Phospho-proteomics, Software, Staining, Irradiation, Two Tailed Test, MANN-WHITNEY