Journal: Cancer medicine

Article Title: Direct effect of dasatinib on signal transduction pathways associated with a rapid mobilization of cytotoxic lymphocytes.

doi: 10.1002/cam4.925

Figure Lengend Snippet: Figure 1. Flow cytometric analysis of each lymphocyte subset and expression of phosphorylated proteins. Representative data are shown. Lymphocyte fractions are classified according to surface antibodies including CD3, CD8, and CD56. Natural killer (NK) cells were defined as the CD3-CD56+ immunophenotype and cytotoxic T lymphocytes (CTLs) were CD3+ CD8+ (A). Cells were costained with phospho-specific antibodies, including antibodies targeting pJAK1, pJAK2, pSTAT1, pSTAT3, pERK, pJNK, pp38, and pAKT, and expression levels of the isotype control and phosphorylated proteins in NK cells are presented (B). The values for the isotype control and each phosphorylated protein are shown as the median fluorescence intensity (MFI).

Article Snippet: Fluorescein isothiocyanate (FITC)- labeled polyclonal rabbit IgG antibodies for phosphorylated proteins, including pJAK1 (Y1034, #3238R), pJAK2 (Y1007, #2485R), pSTAT1 (Y701, #1657R), pSTAT3 (Y705, #1658R), pERK (T202/Y204, #1646R), pJNK (T183/Y185, #1640R), pp38 (T180/Y182, #2210R), pAKT (Y315, #5193R), and isotype control (#0295P) were purchased from Bioss (Wobun, MA, USA).

Techniques: Expressing, Control, Fluorescence

Journal: Cancer medicine

Article Title: Direct effect of dasatinib on signal transduction pathways associated with a rapid mobilization of cytotoxic lymphocytes.

doi: 10.1002/cam4.925

Figure Lengend Snippet: Figure 3. Constitutive levels of phosphorylated proteins including pJAK1, pJAK2, pSTAT1, pSTAT3, pERK, pJNK, pp38, and pAKT in natural killer (NK) cells (A) and cytotoxic T lymphocytes (CTLs) (B) grouped according to treatment (dasatinib [n = 18] or other TKI [n = 12]) are shown. The values for phosphorylated proteins in each fraction are shown as the median fluorescence intensity (MFI).

Article Snippet: Fluorescein isothiocyanate (FITC)- labeled polyclonal rabbit IgG antibodies for phosphorylated proteins, including pJAK1 (Y1034, #3238R), pJAK2 (Y1007, #2485R), pSTAT1 (Y701, #1657R), pSTAT3 (Y705, #1658R), pERK (T202/Y204, #1646R), pJNK (T183/Y185, #1640R), pp38 (T180/Y182, #2210R), pAKT (Y315, #5193R), and isotype control (#0295P) were purchased from Bioss (Wobun, MA, USA).

Techniques: Fluorescence

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: P orphyromonas gingivalis promotes the progression of oral squamous cell carcinoma by activating the neutrophil chemotaxis in the tumour microenvironment

doi: 10.1007/s00262-022-03348-5

Figure Lengend Snippet: Acquisition of the EMT phenotype was verified by activation of the JAK1/STAT3 signalling pathway through the CXCL2/CXCR2 signalling axis. a Western blot analysis showed that EMT occurred and the JAK1/STAT3 signalling pathway was activated by CXCL2/CXCR2 when P. gingivalis infected the TME of OSCC. b After coculture for 24 h, and the cells were transinfected with sh-CXCL2 and sh-CXCR2 lentiviruses for 72 h, the cells were observed under a fluorescence microscope. c, d Knockdown of CXCL2 and CXCR2 was verified at the protein and mRNA levels by Western blotting and qRT-PCR

Article Snippet: Primary antibodies against P. gingivalis (Dia-An, lnc, it was freshly prepared, 1:1000), CXCL2 (bs-1162R, BIOSS, Biotech, Beijing, China, 1:2000), CXCR2 (bs-1629R, BIOSS, Biotech, Beijing, China, 1:2000), JAK1 (bs-1439R, BIOSS, Biotech, Beijing, China, 1:2000), STAT3 (bs-55208R, BIOSS, Biotech, Beijing, China, 1:2000), p-JAK1 (bs-3238R, BIOSS, Biotech, Beijing, China, 1:2000), p-STAT3 (bs-1658R, BIOSS, Biotech, Beijing, China, 1:2000), E-cadherin (bs-10009R, BIOSS, Biotech, Beijing, China, 1:2000), N-cadherin (bs-1172R, BIOSS, Biotech, Beijing, China, 1:2000), and GAPDH (bs-13282R, BIOSS, Biotech, Beijing, China, 1:2000) and secondary antibodies (anti‐rabbit‐HRP, Cell Signalling Technology, Inc, Boston, USA) were used in this study.

Techniques: Activation Assay, Western Blot, Infection, Fluorescence, Microscopy, Knockdown, Quantitative RT-PCR

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: P orphyromonas gingivalis promotes the progression of oral squamous cell carcinoma by activating the neutrophil chemotaxis in the tumour microenvironment

doi: 10.1007/s00262-022-03348-5

Figure Lengend Snippet: Screening and culture of positive clone cells expressing both the lentiviruses of sh-CXCL2 and sh-CXCR2 which can reverse the phenotype of EMT and rescue the signalling pathway of JAK1/STAT3. a The lentiviruses of sh-CXCL2 and sh-CXCR2 were co-transfected into TSCCa cells. b Flow cytometry was used to screen out the positive cells that expression both lentiviruses of sh-CXCL2 and sh-CXCR2. c The clone of positive cells were screened out and cultured by limited dilution method. d Western blot analysis after inactivation of the CXCL2/CXCR2 signalling axis in the P. gingivalis-infected TME of OSCC, the activity of the JAK1/STAT3 signalling pathway was decreased, and the EMT process was reversed. (*p < 0.05, **p < 0.01, and ***p < 0.001; one‐way analysis of variance)

Article Snippet: Primary antibodies against P. gingivalis (Dia-An, lnc, it was freshly prepared, 1:1000), CXCL2 (bs-1162R, BIOSS, Biotech, Beijing, China, 1:2000), CXCR2 (bs-1629R, BIOSS, Biotech, Beijing, China, 1:2000), JAK1 (bs-1439R, BIOSS, Biotech, Beijing, China, 1:2000), STAT3 (bs-55208R, BIOSS, Biotech, Beijing, China, 1:2000), p-JAK1 (bs-3238R, BIOSS, Biotech, Beijing, China, 1:2000), p-STAT3 (bs-1658R, BIOSS, Biotech, Beijing, China, 1:2000), E-cadherin (bs-10009R, BIOSS, Biotech, Beijing, China, 1:2000), N-cadherin (bs-1172R, BIOSS, Biotech, Beijing, China, 1:2000), and GAPDH (bs-13282R, BIOSS, Biotech, Beijing, China, 1:2000) and secondary antibodies (anti‐rabbit‐HRP, Cell Signalling Technology, Inc, Boston, USA) were used in this study.

Techniques: Expressing, Transfection, Flow Cytometry, Cell Culture, Western Blot, Infection, Activity Assay