Journal: EBioMedicine

Article Title: UVB induces cutaneous squamous cell carcinoma progression by de novo ID4 methylation via methylation regulating enzymes

doi: 10.1016/j.ebiom.2020.102835

Figure Lengend Snippet: Sequences of primers used in qPCR.

Article Snippet: The slides were first blocked with normal goat serum at room temperature for 30 min to minimize nonspecific staining, then incubated overnight with primary antibodies against DNMT1 (1:100, Absin Bioscience Inc., Shanghai, China), DNMT3A (1:50, CST, USA), DNMT3B (1:50, Abcam, UK), TET1 (1:100, Absin Bioscience Inc., Shanghai, China), TET2 (1:100, Absin Bioscience Inc., Shanghai, China), and TET3 (1:100, Abcam, UK).

Techniques: Sequencing

Journal: EBioMedicine

Article Title: UVB induces cutaneous squamous cell carcinoma progression by de novo ID4 methylation via methylation regulating enzymes

doi: 10.1016/j.ebiom.2020.102835

Figure Lengend Snippet: Altered expression of DNMTs and TETs in CSCCs compared with adjacent normal tissues. (a) The relative expression levels of DNMTs and TETs in CSCC and adjacent normal tissues. Data are shown as means ± SD. The statistical significance was assessed by the Student's t -test. (b) The Pearson's correlation between expression level (2 −△Ct ) or methylation level of ID4 and expression levels (2 −△△Ct ) of DNMT1, DNMT3A, DNMT3B, TET1, TET2, and TET3, which were measured by MSP and qPCR in the same patient sets.

Article Snippet: The slides were first blocked with normal goat serum at room temperature for 30 min to minimize nonspecific staining, then incubated overnight with primary antibodies against DNMT1 (1:100, Absin Bioscience Inc., Shanghai, China), DNMT3A (1:50, CST, USA), DNMT3B (1:50, Abcam, UK), TET1 (1:100, Absin Bioscience Inc., Shanghai, China), TET2 (1:100, Absin Bioscience Inc., Shanghai, China), and TET3 (1:100, Abcam, UK).

Techniques: Expressing, Methylation

Journal: Frontiers in Pharmacology

Article Title: Lysophosphatidic acid suppresses apoptosis of high-grade serous ovarian cancer cells by inducing autophagy activity and promotes cell-cycle progression via EGFR-PI3K/Aurora-A Thr288 -geminin dual signaling pathways

doi: 10.3389/fphar.2022.1046269

Figure Lengend Snippet: LPA potentiates geminin stability by targeting Aurora-A Thr288 in HGSOC cells. (A,B) Immunoblotting analysis of geminin protein levels in A2780 cells (A) and OVCAR5 cells (B) after pretreated with or without CCT137690 (25 μM) for 120 min, and then stimulated with 0.1% DMSO (vehicle) or 10 μM LPA. Mean ± SEM, n = 3, VNOVA, Dunnett’s Multiple Comparison Test, * p < 0.05, ** p < 0.01, *** p < 0.001. (C) IB analysis of Aurora-A protein levels in A2780 cells and OVCAR5 cells. Mean ± SEM, n = 3, VNOVA, Dunnett’s Multiple Comparison Test, ns, non-significant. (D,E) Immunoprecipitation analysis with anti-phospho-Aurora Kinase (Thr288) antibodies in A2780 cells (D) and OVCAR5 cells (E) treated with 0.1% DMSO (vehicle), 10 μM LPA or 10 ng/mL EGF for 5 min (A2780 cells) or 30 min (OVCAR5 cells). (F) Correlation of gene AURKA and EGFR in TCGA ovarian cancer cohort. log2 fold changes of gene expression on the axis, and the Spearman Method was used for calculating the correlation coefficient. (G) The protein-protein associations between Aurora-A and EGFR protein. Data from STRING. (H–K) IP analysis with anti-phospho-Aurora Kinase (Thr288) antibodies in A2780 cells (H,J) and OVCAR5 cells (I,K) after pretreated with or without 10 μM BB94 (H,I) or 10 μM AG1478 (J,K) for 30 min, then treated with 0.1% DMSO (vehicle), 10 μM LPA or 10 ng/mL EGF for 5 min (A2780 cells) or 30 min (OVCAR5 cells).

Article Snippet: Antibodies were from the following suppliers: anti-phosphotyrosine (EPR16871; Abcam); anti-GAPDH (HC301), anti-β-actin (HC201), anti-β-tubulin (HC101), anti-rabbit (HS101), and anti-mouse (HS201) antibodies (TransGen Biotech, Beijing, China); anti-Bax (bs0127R), anti-Bcl 2 (bs0032R) antibodies (Bioss, Beijing, China); anti-BrdU (abs128684), anti-AURKA (abs136365), anti-phospho-Aurora kinase (Thr288) (abs130639) antibodies (Absin, Shanghai, China); anti-caspase 3/p17 (19677-1-AP), anti-caspase 6/p18/p11 (10198-1-AP), anti-caspase7 (27155-1-AP), anti-caspase 9 (10380-1-AP), anti-PARP (13371-1-AP), anti-EGFR (18986-1-AP), anti-geminin (10802-1-AP) antibodies (Proteintech Group, Wuhan, Hubei, China); anti-LC3B (AF4650, Affinity Biosciences, United States); Alexa Fluor 488–conjugated goat anti-mouse IgG (H + L) antibody (A11008, Invitrogen).

Techniques: Western Blot, Comparison, Immunoprecipitation, Gene Expression

Journal: Cells

Article Title: CD80-Mediated T-Cell Suppression by Cancer Stem-like Cells in Head and Neck Squamous Cell Carcinoma

doi: 10.3390/cells15030266

Figure Lengend Snippet: Anti-CD80 antibodies reinforce T-cell killing ability. ( A ) Calcein AM/PI staining in HNSCC cells after co-culture with or without T cells/anti-CD80 antibodies for 48 h. Scale bar, 200 μm. ( B ) Representative images and quantification of flow cytometry show the dead cell rate of HNSCC cells after co-culture with or without T cells/anti-CD80 antibodies for 48 h. Values are mean ± SD. ns, no significance; * p < 0.05; *** p < 0.001; One-way ANOVA test, n = 3. ( C ) Representative images of flow cytometry show the rate of caspase3 + HNSCC cells after co-culture with or without T cells/anti-CD80 antibodies for 48 h.

Article Snippet: The primary antibodies used were anti-caspase3 antibody (1:100, Absin Bioscience Inc., Shanghai, China, Cat#abs119676), anti-CD8 antibody (1:100, Santa Cruz Biotechnology, Dallas, TX, USA, Cat#sc-1177), anti-GZMB antibody (1:100, Proteintech, Wuhan, China, Cat# 13588-1-AP), anti-perforin antibody (1:100, Proteintech, Wuhan, China, Cat# 14580-1-AP), and anti-IFN- γ antibody (1:100, Proteintech, Wuhan, China, Cat#15365-1-AP).

Techniques: Staining, Co-Culture Assay, Flow Cytometry

Journal: Cells

Article Title: CD80-Mediated T-Cell Suppression by Cancer Stem-like Cells in Head and Neck Squamous Cell Carcinoma

doi: 10.3390/cells15030266

Figure Lengend Snippet: Blockade of CD80 inhibits HNSCC progression and enhances CD8 + T-cell function. ( A ) MOC1 and MOC2 cells were injected subcutaneously into C57BL/6 mice. Representative images show the tumors 18 days after implantation. ( B ) Tumor volume and tumor weight were significantly reduced in C57BL/6 mice treated with anti-CD80 antibody. Values are mean ± SD. ** p < 0.01; *** p < 0.001; Student’s t test; n = 6. ( C ) Representative H&E staining of tumors from mice injected with MOC2 cells. Scale bar, 25 μm. ( D – F ) Representative immunohistochemical staining and quantitative analysis of Caspase3, CD8, GZMB, Perforin, and IFN-γ from MOC2-bearing mice treated with isotype IgG or anti-CD80 antibody. Scale bar, 40 μm. Values are mean ± SD. ** p < 0.01; *** p < 0.001; Student’s t test; n = 6.

Article Snippet: The primary antibodies used were anti-caspase3 antibody (1:100, Absin Bioscience Inc., Shanghai, China, Cat#abs119676), anti-CD8 antibody (1:100, Santa Cruz Biotechnology, Dallas, TX, USA, Cat#sc-1177), anti-GZMB antibody (1:100, Proteintech, Wuhan, China, Cat# 13588-1-AP), anti-perforin antibody (1:100, Proteintech, Wuhan, China, Cat# 14580-1-AP), and anti-IFN- γ antibody (1:100, Proteintech, Wuhan, China, Cat#15365-1-AP).

Techniques: Cell Function Assay, Injection, Staining, Immunohistochemical staining