Journal: Cancer Immunology, Immunotherapy : CII

Article Title: The glucocorticoids prednisone and dexamethasone differentially modulate T cell function in response to anti-PD-1 and anti-CTLA-4 immune checkpoint blockade

doi: 10.1007/s00262-020-02555-2

Figure Lengend Snippet: Dexamethasone treatment impacts the expression of LAG-3 by CD8+ T cells. PBMCs were treated with the indicated concentrations of dexamethasone (dexa), anti-PD-1 (pembrolizumab (pem)), a-CTLA-4 (ipilimumab (ipi)) alone or in combination and stimulated with SEB for 72 h. a Representative FACS plots showing expression of LAG-3 by gated CD8+ T cells in response to dexamethasone treatment and SEB stimulation. b Cumulative data of LAG-3 expression by CD8+ T cells. Data from four independent experiments shown. Error bars represent mean ± SEM. P values obtained by ordinary one-way ANOVA followed by Dunnett’s multiple comparisons test. Unstim, unstimulated cells. c Plots showing phosphorylation of the SHP-2 tyrosine phosphatase Y542 by PD-1 + Jurkat cells stimulated with CHO cells which constitutively express PD-L + and a TCR activator. d PD-1negJurkat cells were pre-treated with the indicated steroids or anti-PD-1 (pembrolizumab) alone or in combination for 48 h. Jurkat cells were co-cultured with CHO cells for 3 h at a ratio of 10:1. Cell lysates were collected followed by measurement of Y542 phosphorylation by ELISA. e PBMCs were pre-treated with the indicated steroids or anti-PD-1 (pembrolizumab) alone or in combination for 3 h. Cell lysates were collected followed by measurement of Y542 phosphorylation by ELISA. Data from four experiments shown. Error bars represent mean ± SD. P values obtained by ordinary one-way ANOVA followed by Dunnett’s multiple comparisons test

Article Snippet: Recombinant Jurkat T cells expressing firefly luciferase gene under the control of NFAT with constitutive expression of PD-1 and Chinese Hamster Ovary (CHO) cells constitutively expressing PD-L1 and an engineered TCR activator were purchased from BPS Bioscience.

Techniques: Expressing, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: The FASEB Journal

Article Title: Deficient transient receptor potential vanilloid type 4 function contributes to compromised [Ca 2+ ] homeostasis in human autosomal‐dominant polycystic kidney disease cells

doi: 10.1096/fj.201701535rr

Figure Lengend Snippet: Figure 9. TRPV4 deglycosylation is associated with reduced channel activity. A) Summary graph of whole-cell TRPV4- dependent current-voltage (I-V) relations upon channel over- expression in CHO cells in control and after pretreatment with tunicamycin (5 mg/ml) for 24 h to block glycosylation. Currents were induced by the application of hypotonic (220 mOsm) medium from isotonic control values (300 mOsm). B) Representative Western blot of whole-cell lysates in CHO cells that overexpress TRPV4 in control and after tunicamycin treatment. Each line represents individual transfection. Lysates were probed with anti-TRPV4 and anti–b-actin Abs. g, glycosy- lated form of TRPV4.

Article Snippet: TRPV4 expression in Chinese hamster ovary cells Chinese hamster ovary (CHO) cells were obtained from American Type Culture Collection (Manassas, VA, USA).

Techniques: Activity Assay, Over Expression, Control, Blocking Assay, Glycoproteomics, Western Blot, Transfection