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a Schematic diagram of I-SceI-induced NHEJ repair in EJ5-GFP U2OS cells. b NHEJ repair efficiency (left) in response to CRTC2 overexpression, and western blot analysis of I-SceI and CRTC2 expression (right) in EJ5-GFP U2OS cells ( n = 3 biologically independent experiments). c NHEJ repair efficiency (left) in response to CRTC2 knockdown, and western blot analysis of I-SceI and CRTC2 expression (right) in EJ5-GFP U2OS cells ( n = 3 biologically independent experiments). Immunofluorescence staining ( d ) and quantitative analysis ( e ) of 53BP1 foci formation at 0, 0.5, 2, 4, 8, and 12 h after IR (6 Gy) in control and CRTC2-knockout HCCLM3 cells ( n = 40 cells per group). Scale bars, 10 µm. f Schematic diagram of VDJ rearrangement and primer locations in pro-B cells. g Pro-B cell sorting process with CD19, B220, IgM, and CD43 from spleen of Crtc2 +/+ and Crtc2 −/− mice (left) and mRNA level of distal and proximal VDJ rearrangement detection by qPCR (right) ( n = 3 mice per group). h Schematic diagram of mouse immunoglobulin class switch recombination (CSR) in mature B cells. i Serum IgM, IgG1, IgG2a, IgG2b, IgG3, and IgA levels in Crtc2 +/+ and Crtc2 −/− mice ( n = 10 mice per group). j Splenic B cells cultured under LPS or <t>LPS/IL4</t> stimulation following CSR to IgG3 (left) and IgE (right) ( n = 3 mice per group). All data are shown as the mean ± SEM. Comparisons between two groups were analyzed by Student’s t test. Comparisons between three or more groups with two or more conditions were assessed by two-way ANOVA. All statistical tests were conducted as two-sided. are provided with this paper.
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a Schematic diagram of I-SceI-induced NHEJ repair in EJ5-GFP U2OS cells. b NHEJ repair efficiency (left) in response to CRTC2 overexpression, and western blot analysis of I-SceI and CRTC2 expression (right) in EJ5-GFP U2OS cells ( n = 3 biologically independent experiments). c NHEJ repair efficiency (left) in response to CRTC2 knockdown, and western blot analysis of I-SceI and CRTC2 expression (right) in EJ5-GFP U2OS cells ( n = 3 biologically independent experiments). Immunofluorescence staining ( d ) and quantitative analysis ( e ) of 53BP1 foci formation at 0, 0.5, 2, 4, 8, and 12 h after IR (6 Gy) in control and CRTC2-knockout HCCLM3 cells ( n = 40 cells per group). Scale bars, 10 µm. f Schematic diagram of VDJ rearrangement and primer locations in pro-B cells. g Pro-B cell sorting process with CD19, B220, IgM, and CD43 from spleen of Crtc2 +/+ and Crtc2 −/− mice (left) and mRNA level of distal and proximal VDJ rearrangement detection by qPCR (right) ( n = 3 mice per group). h Schematic diagram of mouse immunoglobulin class switch recombination (CSR) in mature B cells. i Serum IgM, IgG1, IgG2a, IgG2b, IgG3, and IgA levels in Crtc2 +/+ and Crtc2 −/− mice ( n = 10 mice per group). j Splenic B cells cultured under LPS or <t>LPS/IL4</t> stimulation following CSR to IgG3 (left) and IgE (right) ( n = 3 mice per group). All data are shown as the mean ± SEM. Comparisons between two groups were analyzed by Student’s t test. Comparisons between three or more groups with two or more conditions were assessed by two-way ANOVA. All statistical tests were conducted as two-sided. are provided with this paper.
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Image Search Results


a Schematic diagram of I-SceI-induced NHEJ repair in EJ5-GFP U2OS cells. b NHEJ repair efficiency (left) in response to CRTC2 overexpression, and western blot analysis of I-SceI and CRTC2 expression (right) in EJ5-GFP U2OS cells ( n = 3 biologically independent experiments). c NHEJ repair efficiency (left) in response to CRTC2 knockdown, and western blot analysis of I-SceI and CRTC2 expression (right) in EJ5-GFP U2OS cells ( n = 3 biologically independent experiments). Immunofluorescence staining ( d ) and quantitative analysis ( e ) of 53BP1 foci formation at 0, 0.5, 2, 4, 8, and 12 h after IR (6 Gy) in control and CRTC2-knockout HCCLM3 cells ( n = 40 cells per group). Scale bars, 10 µm. f Schematic diagram of VDJ rearrangement and primer locations in pro-B cells. g Pro-B cell sorting process with CD19, B220, IgM, and CD43 from spleen of Crtc2 +/+ and Crtc2 −/− mice (left) and mRNA level of distal and proximal VDJ rearrangement detection by qPCR (right) ( n = 3 mice per group). h Schematic diagram of mouse immunoglobulin class switch recombination (CSR) in mature B cells. i Serum IgM, IgG1, IgG2a, IgG2b, IgG3, and IgA levels in Crtc2 +/+ and Crtc2 −/− mice ( n = 10 mice per group). j Splenic B cells cultured under LPS or LPS/IL4 stimulation following CSR to IgG3 (left) and IgE (right) ( n = 3 mice per group). All data are shown as the mean ± SEM. Comparisons between two groups were analyzed by Student’s t test. Comparisons between three or more groups with two or more conditions were assessed by two-way ANOVA. All statistical tests were conducted as two-sided. are provided with this paper.

Journal: Nature Communications

Article Title: DNA-PK-mediated CRTC2 phosphorylation promotes NHEJ and suppresses antitumor immunity via relocation to repair complexes

doi: 10.1038/s41467-026-73228-4

Figure Lengend Snippet: a Schematic diagram of I-SceI-induced NHEJ repair in EJ5-GFP U2OS cells. b NHEJ repair efficiency (left) in response to CRTC2 overexpression, and western blot analysis of I-SceI and CRTC2 expression (right) in EJ5-GFP U2OS cells ( n = 3 biologically independent experiments). c NHEJ repair efficiency (left) in response to CRTC2 knockdown, and western blot analysis of I-SceI and CRTC2 expression (right) in EJ5-GFP U2OS cells ( n = 3 biologically independent experiments). Immunofluorescence staining ( d ) and quantitative analysis ( e ) of 53BP1 foci formation at 0, 0.5, 2, 4, 8, and 12 h after IR (6 Gy) in control and CRTC2-knockout HCCLM3 cells ( n = 40 cells per group). Scale bars, 10 µm. f Schematic diagram of VDJ rearrangement and primer locations in pro-B cells. g Pro-B cell sorting process with CD19, B220, IgM, and CD43 from spleen of Crtc2 +/+ and Crtc2 −/− mice (left) and mRNA level of distal and proximal VDJ rearrangement detection by qPCR (right) ( n = 3 mice per group). h Schematic diagram of mouse immunoglobulin class switch recombination (CSR) in mature B cells. i Serum IgM, IgG1, IgG2a, IgG2b, IgG3, and IgA levels in Crtc2 +/+ and Crtc2 −/− mice ( n = 10 mice per group). j Splenic B cells cultured under LPS or LPS/IL4 stimulation following CSR to IgG3 (left) and IgE (right) ( n = 3 mice per group). All data are shown as the mean ± SEM. Comparisons between two groups were analyzed by Student’s t test. Comparisons between three or more groups with two or more conditions were assessed by two-way ANOVA. All statistical tests were conducted as two-sided. are provided with this paper.

Article Snippet: To induce class switching in mouse B cells, they were cultured for 96 h in medium supplemented with LPS (37.5 μg/ml, Solarbio, Cat# 8880) or a mixture of IL4 (10 ng/ml, MCE, Cat# HY-P7080) and LPS 4, 8.

Techniques: Over Expression, Western Blot, Expressing, Knockdown, Immunofluorescence, Staining, Control, Knock-Out, FACS, Cell Culture