Journal: eLife

Article Title: Integrin β4 promotes DNA damage-related drug resistance in triple-negative breast cancer via TNFAIP2/IQGAP1/RAC1

doi: 10.7554/eLife.88483

Figure Lengend Snippet: ( A ) TNFAIP2 interacts with ITGB4.HCC1806 cells with stable TNFAIP2 overexpression were collected from flag-tagged TNFAIP2 cell lysates for immunoprecipitation assays using Flag-M2 beads, and ITGB4 was detected by western blotting (WB). ( B ) Endogenous TNFAIP2 interacts with ITGB4 in HCC1806 cells. Endogenous TNFAIP2 protein was immunoprecipitated using an anti-TNFAIP2 antibody. IgG served as the negative control. Endogenous ITGB4 was detected by WB. ( C ) Endogenous ITGB4 interacts with TNFAIP2 in HCC1806 cells. Endogenous ITGB4 protein was immunoprecipitated using an anti-ITGB4 antibody. IgG served as the negative control. Endogenous TNFAIP2 was detected by WB. ( D–I ) ITGB4 knockdown in HCC1806 and HCC1937 cells significantly decreased cell viability in the presence of EPI (0–800 nM) and BMN (0–40 μM), as measured by the SRB assay. Statistical analysis was performed using one-way analysis of variance (ANOVA), n = 3, *p < 0.05, **p < 0.01, ***p < 0.001. ITGB4 protein expression was detected by WB. ( J–N ) ITGB4 depletion promotes HCC1806 breast cancer cell sensitivity to EPI and BMN treatment in vivo. HCC1806 cells with stable ITGB4 knockdown were transplanted into the fat pad of 7-week-old female nude mice. When the average tumor size reached approximately 50 mm 3 after inoculation, the mice in each group were randomly divided into two subgroups ( n = 4/group) to receive EPI (2.5 mg/kg), BMN (1 mg/kg), or vehicle control for 22 days ( J ). Tumor masses were collected and weighed at the end of the experiments ( K ), and tumor size was measured twice a week ( L–N ). *p < 0.05, **p < 0.01, ***p < 0.001, t -test. ( O ) ITGB4 knockdown increased DNA damage of EPI and BMN. HCC1806 and HCC1937 cells with ITGB4 knockdown were treated with 400 nM EPI for 24 hr and 5 μM BMN for 24 hr, respectively. ITGB4, γH2AX, and PARP protein expression was detected by WB. Figure 5—source data 1. Uncropped western blot images for .

Article Snippet: After 12 hr, the slides were washed three times with PBS and incubated with secondary antibodies hypersensitive enzyme-labeled goat anti-mouse/rabbit IgG polymer (OriGene, China) at room temperature for 20 min, DAB concentrate chromogenic solution (1:200dilution of concentrated DAB chromogenic solution), counterstained with 0.5% hematoxylin, dehydrated with graded concentrations of ethanol for 3 min each (70–80–90–100%), and finally stained with dimethyl benzene immunostained slides were evaluated by light microscopy.

Techniques: Over Expression, Immunoprecipitation, Western Blot, Negative Control, Sulforhodamine B Assay, Expressing, In Vivo

Journal: Frontiers in Immunology

Article Title: The immune checkpoint adenosine 2A receptor is associated with aggressive clinical outcomes and reflects an immunosuppressive tumor microenvironment in human breast cancer

doi: 10.3389/fimmu.2023.1201632

Figure Lengend Snippet: A2AR expression on breast tumors and matched control tissues. The A2AR expression level was measured by qRT-PCR and immunohistochemistry. (A) A2AR gene expression exhibits an elevated level in breast tumors compared to matched control tissues ( p = 0.0176). (C) Representative immunohistochemical staining for A2AR and the IgG2a isotype (magnification20X, scale bar 200µm) in positive control tissues: Placenta (black arrows indicate tubular epithelial lining cells) and Testis (black arrows indicate germline cells at different development stages, and red arrows show Leydig cells). (D) A2AR staining showed membrane and cytoplasmic localization within both tumor and immune cells (black arrows indicate tumor cells, and red arrows show immune cells). (B, D) A2AR protein expression is more pronounced within tumors compared to matched control tissues ( p = 0.0020). Significance was calculated using the Wilcoxon matched-pairs signed rank test. *p<0.05, **p<0.01.

Article Snippet: Negative control sections were incubated with Isotype control antibodies (Mouse IgG2a Isotype Control clone PPV-04 (OriGene) at a 1:500 dilution and Mouse IgG1 Isotype Control clone MOPC-21 (LSBio) at a 1:200 dilution for each sample.

Techniques: Expressing, Quantitative RT-PCR, Immunohistochemistry, Staining, Positive Control, Membrane

Journal: International Journal of Molecular Sciences

Article Title: Administration of Recombinant TAPBPL Protein Ameliorates Collagen-Induced Arthritis in Mice

doi: 10.3390/ijms241813772

Figure Lengend Snippet: TAPBPL reduces CII-specific IgG1, IgG2a, and IgG2b antibody production in CIA mice. CIA-induced DBA/1 mice received control Ig or TAPBPL-Ig protein as in . The blood was collected on day 58. Analysis of mouse serum for anti-CII IgG1, IgG2a, and IgG2b antibody production via ELISA. Results are given as mean ± SD from 1 of 3 similar independent experiments (5–6 mice/group/experiment). * p < 0.05 versus control Ig group.

Article Snippet: We cloned the extracellular domain of the human TAPBPL gene into a pCMV6-AC-FC-S expression vector that contains the constant region of mouse IgG2a (ORIGENE, Rockville, MD, USA).

Techniques: Enzyme-linked Immunosorbent Assay