Journal: Frontiers in Oncology

Article Title: Increased TIGIT expression correlates with impaired NK cell function in diffuse large B-cell lymphoma

doi: 10.3389/fonc.2025.1551061

Figure Lengend Snippet: Enhanced killing ability of NK cells was observed by blocking TIGIT (A) Schematic representation of purity of peripheral NK cells before and after sorting. (B) Comparisons of the killing ability of NK cells towards DLBCL cell line SU-DHL-4 at E:T ratios of 5:1, 10:1 and 20:1 among DLBCL patients (n=6) and HC (n=6). (C) Comparisons of the killing ability of TIGIT + NK cells and TIGIT - NK cells towards DLBCL cell line SU-DHL-4 at an E:T ratio of 20:1 (n=6). (D) Quantification of the killing ability of NK cells from DLBCL patients in the presence of varying concentrations of anti-TIGIT mAb blockade at an E:T ratio of 20:1 (n=6). (E) Rag2-/- mice were subcutaneously injected with 1 × 10 6 A20 tumor cells on day0. At various time points thereafter, the mice were intraperitoneally administered either anti-TIGIT mAb or IgG as a control. Tumor volume was measured at multiple time points (n=6). Each symbol represents the mean value from three technical replicates of one subject, and the data are shown as mean ± SD. HC, healthy control. DLBCL, diffuse B-cell lymphoma. * indicates P<0.05, ** indicates P<0.01, *** indicates P<0.001, **** indicates P<0.0001, ns indicates no significance. Unpaired Student’s t-test was used (B–D) . Two-way analysis of variance was used (E) .

Article Snippet: A20 cells were obtained from American Type Culture Collection (Manassas, USA).

Techniques: Blocking Assay, Injection, Control

Journal: Molecular cell

Article Title: Liquid Phase Separation of NEMO Induced by Polyubiquitin Chains Activates NF-κB

doi: 10.1016/j.molcel.2022.03.037

Figure Lengend Snippet:

Article Snippet: Rabbit monoclonal anti-A20 , Cell Signaling Technology , Cat# 5630.

Techniques: Western Blot, Immunostaining, Recombinant, Software

Journal: Journal of Cellular and Molecular Medicine

Article Title: A20 plays a critical role in the immunoregulatory function of mesenchymal stem cells

doi: 10.1111/jcmm.12849

Figure Lengend Snippet: Inflammatory cytokines induce A20 expression in MSC s. MSC s ( A ) and C3H/10T1/2 ( B and C ) were treated with 0, 2, 5 and 10 ng/ml IFN ‐γ and TNF ‐α for 24 hrs. A20 mRNA and protein levels were examined by qRT ‐ PCR and Western blot analysis. MSC s ( D ) and C3H/10T1/2 ( E and F ) were treated with 5 ng/ml IFN ‐γ and TNF ‐α for 0, 6, 12 and 24 hrs, and mRNA and protein expression levels were determined by qRT ‐ PCR and Western blot analysis, respectively.

Article Snippet: After 24 hrs, C3H/10T1/2 cells were transduced with lentivirus expressing murine A20 shRNA (shA20 C3 MSCs) or control lentivirus (shCTRL C3 MSCs) in the presence of 10 μg/ml polybrene (Santa Cruz Biotechnology, Dallas, TX, USA) for 6 hrs.

Techniques: Expressing, Quantitative RT-PCR, Western Blot

Journal: Journal of Cellular and Molecular Medicine

Article Title: A20 plays a critical role in the immunoregulatory function of mesenchymal stem cells

doi: 10.1111/jcmm.12849

Figure Lengend Snippet: Morphological and phenotypic characterization of C3 MSC s after A20 knockdown. ( A ) qRT ‐ PCR analysis of A20 mRNA levels with or without A20 knockdown. ( B ) Morphology and ( C ) size of cultured sh CTRL C3 MSC s and shA20 C3 MSC s were analysed by microscopy and flow cytometry. ( D ) Cell surface markers were analysed by flow cytometry, ** P < 0.01. All experiments were repeated three times.

Article Snippet: After 24 hrs, C3H/10T1/2 cells were transduced with lentivirus expressing murine A20 shRNA (shA20 C3 MSCs) or control lentivirus (shCTRL C3 MSCs) in the presence of 10 μg/ml polybrene (Santa Cruz Biotechnology, Dallas, TX, USA) for 6 hrs.

Techniques: Knockdown, Quantitative RT-PCR, Cell Culture, Microscopy, Flow Cytometry

Journal: Journal of Cellular and Molecular Medicine

Article Title: A20 plays a critical role in the immunoregulatory function of mesenchymal stem cells

doi: 10.1111/jcmm.12849

Figure Lengend Snippet: A20 knockdown attenuates immunosuppressive capacity of C3 MSC s in vitro . CFSE ‐labelled CD 3 + T cell was cultured alone ( A ) or co‐cultured with different numbers of C3 MSC s ( B ) in RPMI 1640 complete medium supplemented with PMA (50 ng/ml) and ionomycin (1 μg/ml) for 48 hrs. Cells were subjected to flow cytometry for T cell proliferation as detected by the CFSE signal.

Article Snippet: After 24 hrs, C3H/10T1/2 cells were transduced with lentivirus expressing murine A20 shRNA (shA20 C3 MSCs) or control lentivirus (shCTRL C3 MSCs) in the presence of 10 μg/ml polybrene (Santa Cruz Biotechnology, Dallas, TX, USA) for 6 hrs.

Techniques: Knockdown, In Vitro, Cell Culture, Flow Cytometry

Journal: Journal of Cellular and Molecular Medicine

Article Title: A20 plays a critical role in the immunoregulatory function of mesenchymal stem cells

doi: 10.1111/jcmm.12849

Figure Lengend Snippet: A20 knockdown inhibits tumorigenesis in vivo . ( A ) Representative tumours photographed 13 days after C57 BL /6 mice were injected with PBS , sh CTRL C3 MSC s or shA20 C3 MSC s. ( B ) On day 0, 5 × 10 5 B16‐F0 cells in 100‐μl PBS were injected subcutaneously into the right posterior flanks, with or without co‐injection of sh CTRL C3 MSC s or shA20 C3 MSC s (1 × 10 6 cells). Tumour growth was measured at 2‐day intervals, and tumour volume was calculated. ( C ) Thirteen days later, all mice were killed and tumours were sectioned and weighted. Each treatment group included four mice, and data are representative of three independent experiments, ** P < 0.01.

Article Snippet: After 24 hrs, C3H/10T1/2 cells were transduced with lentivirus expressing murine A20 shRNA (shA20 C3 MSCs) or control lentivirus (shCTRL C3 MSCs) in the presence of 10 μg/ml polybrene (Santa Cruz Biotechnology, Dallas, TX, USA) for 6 hrs.

Techniques: Knockdown, In Vivo, Injection

Journal: Journal of Cellular and Molecular Medicine

Article Title: A20 plays a critical role in the immunoregulatory function of mesenchymal stem cells

doi: 10.1111/jcmm.12849

Figure Lengend Snippet: A20 knockdown showed no significant effect on ICAM ‐1, VCAM ‐1, PD ‐L1, PGE 2 or nitric oxide expression. Sh CTRL C3 MSC s and shA20 C3 MSC s were cultured with or without stimulation with 5 ng/ml IFN ‐γ and TNF ‐α. 12 hrs later, the expression of ( A ) ICAM ‐1, ( B ) VCAM ‐1 and ( C ) PD ‐L1 were analysed by flow cytometry. ( D ) Nitric oxide production was measured by Griess assay. ( E ) PGE 2 production was measured by ELISA .

Article Snippet: After 24 hrs, C3H/10T1/2 cells were transduced with lentivirus expressing murine A20 shRNA (shA20 C3 MSCs) or control lentivirus (shCTRL C3 MSCs) in the presence of 10 μg/ml polybrene (Santa Cruz Biotechnology, Dallas, TX, USA) for 6 hrs.

Techniques: Knockdown, Expressing, Cell Culture, Flow Cytometry, Griess Assay, Enzyme-linked Immunosorbent Assay

Journal: Journal of Cellular and Molecular Medicine

Article Title: A20 plays a critical role in the immunoregulatory function of mesenchymal stem cells

doi: 10.1111/jcmm.12849

Figure Lengend Snippet: A20 inhibits TNF ‐α and promotes IL ‐10 production in C3 MSC s, and the p38/ MAPK pathway is involved in A20‐induced immunomodulation. TNF ‐α ( A ) and IL ‐10 ( B , left) expression was examined with or without stimulation with 5 ng/ml IFN ‐γ and TNF ‐α by qRT ‐ PCR . IL ‐10 protein level was determined by ELISA ( B , right). ( C ) The time courses of p38 phosphorylation in response to 5 ng/ml IFN ‐γ and TNF ‐α was determined by immunoblotting, ** P < 0.01, * P < 0.05.

Article Snippet: After 24 hrs, C3H/10T1/2 cells were transduced with lentivirus expressing murine A20 shRNA (shA20 C3 MSCs) or control lentivirus (shCTRL C3 MSCs) in the presence of 10 μg/ml polybrene (Santa Cruz Biotechnology, Dallas, TX, USA) for 6 hrs.

Techniques: Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Phospho-proteomics, Western Blot

Journal: Cell Death & Disease

Article Title: Gasdermin E promotes translocation of p65 and c-jun into nucleus in keratinocytes for progression of psoriatic skin inflammation

doi: 10.1038/s41419-024-06545-5

Figure Lengend Snippet: A Neutrophil infiltration was evaluated by detecting Ly6G and MPO through immunohistochemistry study. Statistical analysis of mean staining intensity was shown. B Quantitative real-time PCR analysis was performed to determine the mRNA levels of Il17a, Il23a, Tnfa, Il1b, Ccl20, Cxcl1, Cxcl2, S100a8, and S100a9 . Statistical analysis was shown. C The percentage of CD4+T cells (CD3+CD4+), Th1 cells (CD3+CD4+IFN-γ+), and Th17 cells (CD3+CD4+IL-17A+) in spleen was determined by flow cytometry. Statistical analysis of specific cell percentages was shown. Scale bar represents 100 μm in A . ns not significant, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: ELISA kits of S100A8/A9 (KE00177), CCL20 (KE00149), and IL-8 (KE00006) were purchased from Proteintech (Wuhan, China).

Techniques: Immunohistochemistry, Staining, Real-time Polymerase Chain Reaction, Flow Cytometry

Journal: Cell Death & Disease

Article Title: Gasdermin E promotes translocation of p65 and c-jun into nucleus in keratinocytes for progression of psoriatic skin inflammation

doi: 10.1038/s41419-024-06545-5

Figure Lengend Snippet: A Proteins of GSDME FL and GSDME NT were detected by western blotting in HaCaT cells at 12, 24, and 48 h after treatment with or without M5. Statistical analysis of interested protein levels was shown. B HaCaT cells were transfected with lentivirus vector containing either NC shRNA or GSDME shRNA, establishing GSDME knockdown cells (sh GSDME) and control cells (sh NC). Knockdown efficiency was evaluated using western blotting assay. Statistical analysis of GSDME was shown. C , D Immunofluorescence assay was used to detect Ki-67 expression or Edu-positive cells. Statistical analysis of mean immunofluorescence intensity was shown. E Quantitative real-time PCR analysis was performed to determine mRNA levels of Il1b, Ccl20, Cxcl1, Cxcl8, S100a8 , and S100a9 . Statistical analysis was shown. F Proteins of GSDME FL, S100A8, and S100A9 in cell lysate were detected by western blotting assay. Statistical analysis of interested protein levels was shown. G Secretion levels of S100A8/A9, CCL20, and CXCL8 were determined by ELISA assay. Statistical analysis was shown. H S100A8 and S100A9 were detected by immunofluorescence assay in mice skin sections, respectively. Statistical analysis was shown. I Proteins of p-p65, p65, p-c-jun, and c-jun in cell lysate were detected by western blotting assay. Statistical analysis of interested protein levels was shown. J Proteins of p65, c-jun, and GSDME-FL were detected by western blotting assay in lysate of cell plasma or nucleus, respectively. Statistical analysis of interested protein levels was shown. GAPDH and HDAC1 served as a loading control of cell plasma protein and nucleus proteins, respectively. K , L Immunofluorescence assay was used to detect translocation of p65 and c-jun into nucleus. M Co-immunoprecipitation assay was used to detect the interaction between GSDME-FL and p65 or c-jun. Scale bar represents 100 μm in C , D , H , K and L . ns not significant, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: ELISA kits of S100A8/A9 (KE00177), CCL20 (KE00149), and IL-8 (KE00006) were purchased from Proteintech (Wuhan, China).

Techniques: Western Blot, Transfection, Plasmid Preparation, shRNA, Knockdown, Control, Immunofluorescence, Expressing, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Clinical Proteomics, Translocation Assay, Co-Immunoprecipitation Assay

Journal: Cell Death & Disease

Article Title: Gasdermin E promotes translocation of p65 and c-jun into nucleus in keratinocytes for progression of psoriatic skin inflammation

doi: 10.1038/s41419-024-06545-5

Figure Lengend Snippet: Primer sequences in this study.

Article Snippet: ELISA kits of S100A8/A9 (KE00177), CCL20 (KE00149), and IL-8 (KE00006) were purchased from Proteintech (Wuhan, China).

Techniques: