Journal: Cell Death & Disease

Article Title: A HER2-targeting antibody-MMAE conjugate RC48 sensitizes immunotherapy in HER2-positive colon cancer by triggering the cGAS-STING pathway

doi: 10.1038/s41419-023-06073-8

Figure Lengend Snippet: A Analysis of the immunohistochemical pathological reports for HER2 of 13 CRC patients in Jiangsu Provincial Hospital. B Western blot analysis of HER2 expression in murine and human CRC cell lines. C Expression and localization of HER2 detected by immunofluorescence on MC38 HER2 , MC38 Ctrl , CT26 HER2 , and CT26 Ctrl cells. D The mRNA expression of HER2 was measured by qPCR. ** P < 0.01. E The expressions of HER2 on the cell membrane were detected using flow cytometry. F The expression and localization of HER2 in HCT116 and HT29 cells were determined by immunofluorescence. Scale bars, 10 μm.

Article Snippet: Anti-HER2 antibody (ab134182) was from Abcam plc.

Techniques: Immunohistochemical staining, Western Blot, Expressing, Immunofluorescence, Membrane, Flow Cytometry

Journal: Cell Death & Disease

Article Title: A HER2-targeting antibody-MMAE conjugate RC48 sensitizes immunotherapy in HER2-positive colon cancer by triggering the cGAS-STING pathway

doi: 10.1038/s41419-023-06073-8

Figure Lengend Snippet: A In vitro growth inhibition assay for HCT116 cells treated with MMAE and Paclitaxel for 24 h. B In vitro growth inhibition assay for HCT116 and HT29 cells. C In vitro growth inhibition assay for MC38 HER2 and MC38 Ctrl cells treated with RC48 for 48 h. D In vitro growth inhibition assay for MC38 Ctrl and MC38 HER2 cells treated with MMAE for 24 h. E Flow cytometry analysis of cell cycle after treatment with different concentrations of RC48 for 48 h. F Clonogenic assay of HCT116, CT26 HER2 and MC38 HER2 treated with RC48. Data represent the mean ± SEM of three replicates. * P < 0.05, ** P < 0.01, versus as indicated. n.s. not significant.

Article Snippet: Anti-HER2 antibody (ab134182) was from Abcam plc.

Techniques: In Vitro, Growth Inhibition Assay, Flow Cytometry, Clonogenic Assay

Journal: Cell Death & Disease

Article Title: A HER2-targeting antibody-MMAE conjugate RC48 sensitizes immunotherapy in HER2-positive colon cancer by triggering the cGAS-STING pathway

doi: 10.1038/s41419-023-06073-8

Figure Lengend Snippet: A MC38 Ctrl cells were treated with different concentrations of Disitamab (0, 10, 30, 100 μg/ml) for 48 h, and the expression of p-TBK1, p-IRF3, TBK1, IRF3 protein was detected by immunoblotting. B MC38 HER2 and MC38 Ctrl cells were treated with 100 μg/ml RC48 for 48 h and 30 μM CPT11 for 12 h and the expression of p-TBK1, p-IRF3, TBK1, IRF3 protein was detected by immunoblotting. C HCT116 was treated with 100 μg/ml RC48 for 48 h and 30 μM CPT11 for 12 h. D The expressions of HER2 on the cell membrane were detected using flow cytometry. MFI Mean fluorescence intensity. E Co-immunoprecipitation of endogenous HER2 and STING in HCT116. F The expression and localization of HER2 in MC38 HER2 were detected by immunofluorescence. * P < 0.05, ** P < 0.01 versus as indicated. n.s. not significant. Scale bar, 10 μm.

Article Snippet: Anti-HER2 antibody (ab134182) was from Abcam plc.

Techniques: Expressing, Western Blot, Membrane, Flow Cytometry, Fluorescence, Immunoprecipitation, Immunofluorescence

Journal: Cell Death & Disease

Article Title: A HER2-targeting antibody-MMAE conjugate RC48 sensitizes immunotherapy in HER2-positive colon cancer by triggering the cGAS-STING pathway

doi: 10.1038/s41419-023-06073-8

Figure Lengend Snippet: A – E MC38 HER2 (1 × 10 6 ) cells were inoculated in the C57BL/6 mice. Drugs were given as described in Materials and Methods. B , C Tumor volume and body weight were measured every two days. D , E Solid tumors were separated and weighed after the mice were sacrificed. The data represent the mean ± SEM of 10 mice per group. F Paraffin sections of tumor tissues were analyzed by H&E staining, PCNA immunohistochemistry staining, and TUNEL fluorescent staining. G The survival of tumor-bearing mice. H – M Same experiment with CT26 HER2 in BALB/c mice. The data represent the mean ± SEM of 7 mice per group. * P < 0.05, ** P < 0.01 versus as indicated. n.s. not significant. Scale bar, 50 μm.

Article Snippet: Anti-HER2 antibody (ab134182) was from Abcam plc.

Techniques: Staining, Immunohistochemistry, TUNEL Assay

Journal: Cell Death & Disease

Article Title: A HER2-targeting antibody-MMAE conjugate RC48 sensitizes immunotherapy in HER2-positive colon cancer by triggering the cGAS-STING pathway

doi: 10.1038/s41419-023-06073-8

Figure Lengend Snippet: A Immunofluorescence staining for p-TBK1 and p-IRF3 in tumor tissue in C57BL/6. B , C Immunohistochemical detection of IFN-β secretion in tumor tissue in BALB/c and C57BL/6. D LDH release assay was performed using Tils achieved from the syngeneic model of CT26 HER2 as described in material and methods. E , F Examination of CD3 + , CD8 + , IFN-γ secretion in the tumor by flow cytometry in BALB/c and C57BL/6. * P < 0.05, ** P < 0.01 versus as indicated. n.s. not significant. Scale bar, 100 μm.

Article Snippet: Anti-HER2 antibody (ab134182) was from Abcam plc.

Techniques: Immunofluorescence, Staining, Immunohistochemical staining, Lactate Dehydrogenase Assay, Flow Cytometry

Journal: Cell Death & Disease

Article Title: A HER2-targeting antibody-MMAE conjugate RC48 sensitizes immunotherapy in HER2-positive colon cancer by triggering the cGAS-STING pathway

doi: 10.1038/s41419-023-06073-8

Figure Lengend Snippet: RC48 exhibits the capability to bind to HER2 receptors and undergo internalization, subsequently releasing the toxin MMAE. MMAE inhibits tubulin, leading to the disruption of the cell cycle and resulting in the efficient elimination of tumor cells. Moreover, through its interaction with HER2, RC48 can alleviate the inhibition induced by internalized HER2 on the STING pathway and its downstream signaling. These combined mechanisms enable RC48 to effectively impede the proliferation of HER2-positive tumors while also enhancing their sensitivity to immunotherapy.

Article Snippet: Anti-HER2 antibody (ab134182) was from Abcam plc.

Techniques: Disruption, Inhibition

Journal: Annals of Medicine

Article Title: Multi-omics analysis and validation of autophagy-related diagnostic biomarker in osteoarthritis

doi: 10.1080/07853890.2025.2548045

Figure Lengend Snippet: Analysis and validation of signature genes. (A) Violin plot of characteristic gene expression in normal and OA samples. (B) Line plot of signature gene expression in normal and OA samples. (C) ROC curves to validate the diagnostic accuracy of signature genes. (D) Expression of ERBB2 in the GSE48556 dataset.

Article Snippet: Next, they were blocked with 10% goat serum (Gibco, USA) at room temperature for 20 min and then incubated overnight with the primary antibody of ERBB2 (Proteintech, 18299-1-AP, 1:300).

Techniques: Biomarker Discovery, Gene Expression, Diagnostic Assay, Expressing

Journal: Annals of Medicine

Article Title: Multi-omics analysis and validation of autophagy-related diagnostic biomarker in osteoarthritis

doi: 10.1080/07853890.2025.2548045

Figure Lengend Snippet: Analysis of ERBB2 expression. (A) Heat maps of the top 20 DEGs in high and low ERBB2 expression groups. (B) Volcano map of DEGs in ERBB2 high and low expression groups. (D) Heat map of the correlation between ERBB2 and DEGs, with positive correlation in red and negative correlation in blue.

Article Snippet: Next, they were blocked with 10% goat serum (Gibco, USA) at room temperature for 20 min and then incubated overnight with the primary antibody of ERBB2 (Proteintech, 18299-1-AP, 1:300).

Techniques: Expressing

Journal: Annals of Medicine

Article Title: Multi-omics analysis and validation of autophagy-related diagnostic biomarker in osteoarthritis

doi: 10.1080/07853890.2025.2548045

Figure Lengend Snippet: Immune-related function analysis in samples of high and low ERBB2 expression groups (* p < 0.05, ** p < 0.01).

Article Snippet: Next, they were blocked with 10% goat serum (Gibco, USA) at room temperature for 20 min and then incubated overnight with the primary antibody of ERBB2 (Proteintech, 18299-1-AP, 1:300).

Techniques: Expressing

Journal: Annals of Medicine

Article Title: Multi-omics analysis and validation of autophagy-related diagnostic biomarker in osteoarthritis

doi: 10.1080/07853890.2025.2548045

Figure Lengend Snippet: GSEA and GSVA analysis of ERBB2. (A–B) GSEA analysis of ERBB2 in high expression group. (C–D) GSEA analysis of ERBB2 in low expression group. (E) E GSVA analysis of biological processes, cellular components, and molecular functions of ERBB2. (F) Pathway-related GSVA analysis of ERBB2.

Article Snippet: Next, they were blocked with 10% goat serum (Gibco, USA) at room temperature for 20 min and then incubated overnight with the primary antibody of ERBB2 (Proteintech, 18299-1-AP, 1:300).

Techniques: Expressing

Journal: Annals of Medicine

Article Title: Multi-omics analysis and validation of autophagy-related diagnostic biomarker in osteoarthritis

doi: 10.1080/07853890.2025.2548045

Figure Lengend Snippet: ceRNA Regulatory network and potential drug action network. (A) ceRNA network regulation diagram of lncRNA-miRNA-ERBB2. (B) Potential drug-signature gene network diagram of OA, red for ERBB2, blue for drug.

Article Snippet: Next, they were blocked with 10% goat serum (Gibco, USA) at room temperature for 20 min and then incubated overnight with the primary antibody of ERBB2 (Proteintech, 18299-1-AP, 1:300).

Techniques:

Journal: Annals of Medicine

Article Title: Multi-omics analysis and validation of autophagy-related diagnostic biomarker in osteoarthritis

doi: 10.1080/07853890.2025.2548045

Figure Lengend Snippet: ERBB2 Transcription levels and protein expression levels between normal and OA samples. (A) Expression level of ERBB2 in qRT-PCR. (B) Protein expression level of ERBB2. (C) Immunohistochemical results of normal human cartilage and OA cartilage from patients (* p < 0.05; ** p < 0.01; *** p < 0.001).

Article Snippet: Next, they were blocked with 10% goat serum (Gibco, USA) at room temperature for 20 min and then incubated overnight with the primary antibody of ERBB2 (Proteintech, 18299-1-AP, 1:300).

Techniques: Expressing, Quantitative RT-PCR, Immunohistochemical staining

Journal: Annals of Medicine

Article Title: Multi-omics analysis and validation of autophagy-related diagnostic biomarker in osteoarthritis

doi: 10.1080/07853890.2025.2548045

Figure Lengend Snippet: Tissue staining and immunohistochemical analysis. (A) Hematoxylin-eosin staining of normal and OA rat knee joints. (B) so/FG staining of normal and OA rat knee joints. (C) OARSI score. (D-E) Protein expression level and quantitative analysis of ERBB2 in knee cartilage by IHC (* p < 0.05; ** p < 0.01; *** p < 0.001).

Article Snippet: Next, they were blocked with 10% goat serum (Gibco, USA) at room temperature for 20 min and then incubated overnight with the primary antibody of ERBB2 (Proteintech, 18299-1-AP, 1:300).

Techniques: Staining, Immunohistochemical staining, Expressing, Paraffin-embedded Immunohistochemistry