Journal: iScience

Article Title: TRIM36 and CAMK2N2 regulate ferroptosis and antigen presentation in small cell lung cancer

doi: 10.1016/j.isci.2026.115310

Figure Lengend Snippet: Survival analysis of patients based on key genes and their correlation with APM genes (A–D) Survival curves of groups with high and low expression of key genes ( GPX3 , CAMK2N2 , TRIM36 , and RND2 ) (George’s cohort ). (E) Scatterplot of the average expression of key genes vs. the average correlation with antigen presentation genes. (F and G) Scatterplots of the correlation between TRIM36, CAMK2N2 and HLA-A, HLA-B, HLA-C, B2M, respectively. Statistical significance was defined as p < 0.05. Statistical analyses were performed using R software. Survival analysis was conducted using the Kaplan-Meier method, and differences between groups were compared using the log rank test. Correlations were analyzed using Pearson’s correlation analysis. HLA, human leukocyte antigen; B2M, beta-2 microglobulin.

Article Snippet: After dewaxing the sections, they were blocked with goat serum, then incubated with primary antibodies against B2M (Boster: A00456-2, Servicebio: GB21303) and β-catenin (Proteintech: Cat#66379-1-Ig).

Techniques: Expressing, Immunopeptidomics, Software

Journal: iScience

Article Title: TRIM36 and CAMK2N2 regulate ferroptosis and antigen presentation in small cell lung cancer

doi: 10.1016/j.isci.2026.115310

Figure Lengend Snippet: TRIM36 and CAMK2N2 promote tumor growth in vivo , inhibit the expression of MHC I, and suppress CD8 + T cell activation in SCLC (A) qRT-PCR detection of the relative expression levels of Camk2n2 gene in the NC group and Sh Camk2n2 group, and Trim36 gene in the NC group and Sh Trim36 group of RP1 cells. qRT-PCR data were normalized to β-actin (ACTB) expression. (B) Gross morphology of subcutaneous tumors in the NC group, Sh Camk2n2 group, and Sh Trim36 group after dissection on the 24th day when mice were injected with cells ( n = 5 per group). (C) Statistics of tumor weights in each group. (D) Curves of tumor volume changes over time in each group. (E and F) Flow cytometry detection and statistics of the expression of H2D/H2K and B2M in tumors. (G–I) Flow cytometry analysis of the proportion of CD3 + CD8 + T cells and IFNγ + CD8 + T cells. Cell experiments were independently repeated 3 times ( n = 3). Data are presented as mean ± SD. Statistical significance: ∗∗ p < 0.01, ∗∗∗ p < 0.001. Statistical analyses were performed using GraphPad Prism software. Multiple group comparisons were conducted using one-way analysis of variance (one-way ANOVA) followed by Bonferroni correction for pairwise comparisons. NC, negative control; MFI, mean fluorescence intensity.

Article Snippet: After dewaxing the sections, they were blocked with goat serum, then incubated with primary antibodies against B2M (Boster: A00456-2, Servicebio: GB21303) and β-catenin (Proteintech: Cat#66379-1-Ig).

Techniques: In Vivo, Expressing, Activation Assay, Quantitative RT-PCR, Dissection, Injection, Flow Cytometry, Software, Negative Control, Fluorescence

Journal: iScience

Article Title: TRIM36 and CAMK2N2 regulate ferroptosis and antigen presentation in small cell lung cancer

doi: 10.1016/j.isci.2026.115310

Figure Lengend Snippet: TRIM36 and CAMK2N2 inhibit the expression of antigen presentation genes in SCLC cells (A and B) The relative expression levels of H2D , H2K , B2m , and Tap1 genes in the untreated group, NC group, Sh Camk2n2 group, and Sh Trim36 group of RP1 cells with or without IFNγ treatment detected by qRT-PCR. qRT-PCR data were normalized to ACTB expression. (C and D) The relative expression levels of Psmb8 , Psmb9 , and Nlrc5 genes in the untreated group, NC group, Sh Camk2n2 group, and Sh Trim36 group with or without IFNγ treatment detected by qRT-PCR. qRT-PCR data were normalized to ACTB expression. (E and F) WB detection of the expression levels of HLA-ABC, B2M, and ACTB proteins in the untreated group, NC group, Sh CAMK2N2 group, and Sh TRIM36 group of H1688 cells with or without IFNγ treatment. WB band intensities were quantified using ImageJ software (normalized to ACTB). (G–J) MFI of H2D/H2K and B2M in the untreated group, NC group, Sh Camk2n2 group, and Sh Trim36 group with or without IFNγ treatment in RP1 cells detected by flow cytometry. All cell experiments were independently repeated three times ( n = 3). Data are presented as mean ± SD. Statistical significance definitions: n.s., no significance; ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001. Statistical analyses were performed using GraphPad Prism software. Multiple group comparisons were conducted using one-way ANOVA followed by Bonferroni correction. NC, negative control; HLA, human leukocyte antigen; B2M, beta-2 microglobulin; H2D/H2K, histocompatibility 2-D region antigen/H2-K region antigen; MFI, mean fluorescence intensity.

Article Snippet: After dewaxing the sections, they were blocked with goat serum, then incubated with primary antibodies against B2M (Boster: A00456-2, Servicebio: GB21303) and β-catenin (Proteintech: Cat#66379-1-Ig).

Techniques: Expressing, Immunopeptidomics, Quantitative RT-PCR, Software, Flow Cytometry, Negative Control, Fluorescence

Journal: iScience

Article Title: TRIM36 and CAMK2N2 regulate ferroptosis and antigen presentation in small cell lung cancer

doi: 10.1016/j.isci.2026.115310

Figure Lengend Snippet: Survival analysis of patients based on key genes and their correlation with APM genes (A–D) Survival curves of groups with high and low expression of key genes ( GPX3 , CAMK2N2 , TRIM36 , and RND2 ) (George’s cohort ). (E) Scatterplot of the average expression of key genes vs. the average correlation with antigen presentation genes. (F and G) Scatterplots of the correlation between TRIM36, CAMK2N2 and HLA-A, HLA-B, HLA-C, B2M, respectively. Statistical significance was defined as p < 0.05. Statistical analyses were performed using R software. Survival analysis was conducted using the Kaplan-Meier method, and differences between groups were compared using the log rank test. Correlations were analyzed using Pearson’s correlation analysis. HLA, human leukocyte antigen; B2M, beta-2 microglobulin.

Article Snippet: Flow cytometry: Stably transfected RP1 cells were stained with fluorescently labeled antibodies against H2D/H2K (Biolegend: Cat#114615) and B2M (Boster: A00456-2, Servicebio: GB21303) to detect the expression of APM genes on the cell membrane surface.

Techniques: Expressing, Immunopeptidomics, Software

Journal: iScience

Article Title: TRIM36 and CAMK2N2 regulate ferroptosis and antigen presentation in small cell lung cancer

doi: 10.1016/j.isci.2026.115310

Figure Lengend Snippet: TRIM36 and CAMK2N2 promote tumor growth in vivo , inhibit the expression of MHC I, and suppress CD8 + T cell activation in SCLC (A) qRT-PCR detection of the relative expression levels of Camk2n2 gene in the NC group and Sh Camk2n2 group, and Trim36 gene in the NC group and Sh Trim36 group of RP1 cells. qRT-PCR data were normalized to β-actin (ACTB) expression. (B) Gross morphology of subcutaneous tumors in the NC group, Sh Camk2n2 group, and Sh Trim36 group after dissection on the 24th day when mice were injected with cells ( n = 5 per group). (C) Statistics of tumor weights in each group. (D) Curves of tumor volume changes over time in each group. (E and F) Flow cytometry detection and statistics of the expression of H2D/H2K and B2M in tumors. (G–I) Flow cytometry analysis of the proportion of CD3 + CD8 + T cells and IFNγ + CD8 + T cells. Cell experiments were independently repeated 3 times ( n = 3). Data are presented as mean ± SD. Statistical significance: ∗∗ p < 0.01, ∗∗∗ p < 0.001. Statistical analyses were performed using GraphPad Prism software. Multiple group comparisons were conducted using one-way analysis of variance (one-way ANOVA) followed by Bonferroni correction for pairwise comparisons. NC, negative control; MFI, mean fluorescence intensity.

Article Snippet: Flow cytometry: Stably transfected RP1 cells were stained with fluorescently labeled antibodies against H2D/H2K (Biolegend: Cat#114615) and B2M (Boster: A00456-2, Servicebio: GB21303) to detect the expression of APM genes on the cell membrane surface.

Techniques: In Vivo, Expressing, Activation Assay, Quantitative RT-PCR, Dissection, Injection, Flow Cytometry, Software, Negative Control, Fluorescence

Journal: iScience

Article Title: TRIM36 and CAMK2N2 regulate ferroptosis and antigen presentation in small cell lung cancer

doi: 10.1016/j.isci.2026.115310

Figure Lengend Snippet: TRIM36 and CAMK2N2 inhibit the expression of antigen presentation genes in SCLC cells (A and B) The relative expression levels of H2D , H2K , B2m , and Tap1 genes in the untreated group, NC group, Sh Camk2n2 group, and Sh Trim36 group of RP1 cells with or without IFNγ treatment detected by qRT-PCR. qRT-PCR data were normalized to ACTB expression. (C and D) The relative expression levels of Psmb8 , Psmb9 , and Nlrc5 genes in the untreated group, NC group, Sh Camk2n2 group, and Sh Trim36 group with or without IFNγ treatment detected by qRT-PCR. qRT-PCR data were normalized to ACTB expression. (E and F) WB detection of the expression levels of HLA-ABC, B2M, and ACTB proteins in the untreated group, NC group, Sh CAMK2N2 group, and Sh TRIM36 group of H1688 cells with or without IFNγ treatment. WB band intensities were quantified using ImageJ software (normalized to ACTB). (G–J) MFI of H2D/H2K and B2M in the untreated group, NC group, Sh Camk2n2 group, and Sh Trim36 group with or without IFNγ treatment in RP1 cells detected by flow cytometry. All cell experiments were independently repeated three times ( n = 3). Data are presented as mean ± SD. Statistical significance definitions: n.s., no significance; ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001. Statistical analyses were performed using GraphPad Prism software. Multiple group comparisons were conducted using one-way ANOVA followed by Bonferroni correction. NC, negative control; HLA, human leukocyte antigen; B2M, beta-2 microglobulin; H2D/H2K, histocompatibility 2-D region antigen/H2-K region antigen; MFI, mean fluorescence intensity.

Article Snippet: Flow cytometry: Stably transfected RP1 cells were stained with fluorescently labeled antibodies against H2D/H2K (Biolegend: Cat#114615) and B2M (Boster: A00456-2, Servicebio: GB21303) to detect the expression of APM genes on the cell membrane surface.

Techniques: Expressing, Immunopeptidomics, Quantitative RT-PCR, Software, Flow Cytometry, Negative Control, Fluorescence