Journal: Journal of Dental Sciences

Article Title: Overexpression of sprouty 1 protein in human oral squamous cell carcinogenesis

doi: 10.1016/j.jds.2020.07.013

Figure Lengend Snippet: Western blot analyses: phosphor-ERK and total-ERK in human oral cancer cell lines as compared with a primary culture of normal oral mucosa (HOK). Upregulation of phosphor-ERK (normalized to total-ERK) in OECM1 and Ca922 cell lines as compared with HOK, and a slightly decreased expression in SAS as compared with HOK. Results were quantified using densitometric analysis, normalized to the level of β-actin, and expressed as a fold change relative to the normal oral mucosa. Bars represent means ± standard deviation of the mean (∗ P < 0.05). A representative result of three independent experiments is shown.

Article Snippet: Further, samples were analyzed using 10% SDS-PAGE (Sigma-Aldrich) gels, and the proteins were transmitted onto a PVDF membrane (Sigma-Aldrich) using Bio-Rad's transblot with primary antibodies against phosphor-ERK (Boster Biological Technology, CA, USA; Cat. No. P00104; 1:1000) and total-ERK (Boster Biological Technology; Cat. No. P00104; 1:1000), with species specificity for human tissues and an observed molecular weight of 42–44 kDa; and β-actin (Sigma-Aldrich; 1:1000), followed by horseradish peroxidase (HRP)-conjugated secondary antibodies (Sigma-Aldrich; 1:5000).

Techniques: Western Blot, Expressing, Standard Deviation

Journal: Frontiers in Medicine

Article Title: CD107a + (LAMP-1) Cytotoxic CD8 + T-Cells in Lupus Nephritis Patients

doi: 10.3389/fmed.2021.556776

Figure Lengend Snippet: Peripheral circulating CD107a + CD8 + T-cells. (A) The percentages of CD107a + CD8 + T-cells in healthy controls (HC), SLE-patients (SLE), (B) patients with lupus nephritis (with LN) and without lupus nephritis (without LN) are shown. (C) The percentages of CD107a + CD8 + T-cells in healthy controls (HC), active SLE-patients (SLE) and inactive patients are shown. Frequencies of these T-cells are shown. Horizontal lines represent the mean. P -values were calculated using the non-parametric Mann-Whitney U-test. (D) Correlation between percentages of CD107a + CD8 + T-cells and disease activity ( n = 30) as assessed by the systemic lupus erythematosus disease activity index (SLEDAI). Spearman analysis was performed to calculate the correlation. A p -value <0.05 was considered significant. (E) A representative dot plot of the flow-cytometry staining is shown for a healthy control (HC), a patient with lupus nephritis (LN) and without LN. The corresponding isotype control is illustrated.

Article Snippet: Incubation with a monoclonal mouse anti-human CD8 (clone C8/144, DAKO, Carpinteria, USA) or polyclonal rabbit anti-human CD107a (polyclonal, Bio-Rad, Munich, Germany) was performed for 60 min at room temperature.

Techniques: MANN-WHITNEY, Activity Assay, Flow Cytometry, Staining, Control

Journal: Frontiers in Medicine

Article Title: CD107a + (LAMP-1) Cytotoxic CD8 + T-Cells in Lupus Nephritis Patients

doi: 10.3389/fmed.2021.556776

Figure Lengend Snippet: (A) The proportion of CD107a + CD8 + T-cells we were analyzed according to the immunosuppressive treatment. Patients were subgrouped in (i) no treatment or prednisone alone (ii) prednisone and mycophenolate mofetil, azathioprine or cyclosporine (iii) prednisone and hydroxychloroquine (iv) prednisone and hydroxychloroquine combined with mycophenolate mofetil, azathioprine or cyclosporine. Patients were compared to healthy controls (HC). P -values were calculated using the non-parametric Mann-Whitney U-test. (B) Correlation between percentages of CD107a + CD8 + T-cells for all samples taken ( n = 30) daily dose of hydroxychloroquine is shown. Spearman analysis was performed to calculate the correlation. A p -value < 0.05 was considered significant.

Article Snippet: Incubation with a monoclonal mouse anti-human CD8 (clone C8/144, DAKO, Carpinteria, USA) or polyclonal rabbit anti-human CD107a (polyclonal, Bio-Rad, Munich, Germany) was performed for 60 min at room temperature.

Techniques: MANN-WHITNEY

Journal: Frontiers in Medicine

Article Title: CD107a + (LAMP-1) Cytotoxic CD8 + T-Cells in Lupus Nephritis Patients

doi: 10.3389/fmed.2021.556776

Figure Lengend Snippet: CD8 + and CD107a + T-cell infiltrates in lupus nephritis. This figure shows representative immunohistochemical staining with anti-CD8 of a tonsil which served as positive control (A,B) . Immunhistochemical staining of a lupus nephritis renal biopsy shows an overview (C) with one glomerulum and interstitial lymphocytes. Several of these lymphocytes express CD8 as demonstrated in (D) . Next, representative immunohistochemical staining with anti-CD107a of a tonsil which served as positive control (E,F) . Immunohistochemical staining lupus nephritis renal biopsy shows an overview (G) with one glomerulum and interstitial lymphocytes. Several of these lymphocytes express CD107a as demonstrated in (H) .

Article Snippet: Incubation with a monoclonal mouse anti-human CD8 (clone C8/144, DAKO, Carpinteria, USA) or polyclonal rabbit anti-human CD107a (polyclonal, Bio-Rad, Munich, Germany) was performed for 60 min at room temperature.

Techniques: Immunohistochemical staining, Staining, Positive Control

Journal: Frontiers in Medicine

Article Title: CD107a + (LAMP-1) Cytotoxic CD8 + T-Cells in Lupus Nephritis Patients

doi: 10.3389/fmed.2021.556776

Figure Lengend Snippet: CD8 + CD107a + immunofluorescence staining. A staining for CD8 Cy3 (red), CD107a FITC (green) and colocalization of CD8/CD107a/DAPI was performed in a tonsil as positive control (A–C) and a representative renal biopsy of an SLE patient with lupus nephritis (WHO class IV) (D–F) . A magnification of a double-positive CD8 + CD107a + kidney infiltrating cell is shown in (G–J) . All scales represent 100 μm.

Article Snippet: Incubation with a monoclonal mouse anti-human CD8 (clone C8/144, DAKO, Carpinteria, USA) or polyclonal rabbit anti-human CD107a (polyclonal, Bio-Rad, Munich, Germany) was performed for 60 min at room temperature.

Techniques: Immunofluorescence, Staining, Positive Control

Journal: Frontiers in Medicine

Article Title: CD107a + (LAMP-1) Cytotoxic CD8 + T-Cells in Lupus Nephritis Patients

doi: 10.3389/fmed.2021.556776

Figure Lengend Snippet: Renal CD8 + and CD107a + T-cells. Correlation between the CD8 + T-cell count (cells/mm 2 ) in renal biopsies of nine lupus nephritis patients and renal histopathology parameters activity index (AI), chronicity index (CI) and proteinuria (g/d). The same correlation was performed for CD107a + T-cell count (cells/mm 2 ) in renal biopsies. Spearman analysis was performed to calculate the correlation. A p -value < 0.05 was considered significant.

Article Snippet: Incubation with a monoclonal mouse anti-human CD8 (clone C8/144, DAKO, Carpinteria, USA) or polyclonal rabbit anti-human CD107a (polyclonal, Bio-Rad, Munich, Germany) was performed for 60 min at room temperature.

Techniques: Cell Counting, Histopathology, Activity Assay