Extended Data Fig. 9: Pcyt2-overexpression in B16 melanoma does not alter PD1+Tim-3+ frequency among the endogenous CD8+ T cells.
From: Tumour interstitial fluid-enriched phosphoethanolamine suppresses T cell function
A) DAG expression of the tumor infiltrating transferred Pmel T cells from the tumors in Fig. 6a. Bar graph on the left shows normalized DAG MFI ±SEM from three independent experiments (n = 7 mice/group, p = 0.0729) B) Western blot confirmation for Pcty2 and Actin in B16 EV and B16-Pcyt2 melanoma cell lines. (Data show n = 3 lysates from 1 independent experiment) Proteins were probed across multiple independent blots that were loaded with the same original proteins. C) PD1 and Tim3 co-expression of endogenous CD8+ T cells from tumors in Fig. 6i, Data quantified on the left show % PD1hiTim3hi from three independent experiments (n = 14-15 mice/group, p = 0.8162). Statistical significance for all indicated comparisons in this figure was calculated using a two-tailed unpaired t-test. D) Data from Fig. 6k were analyzed for Normalized TNF MFI of Live CD8+ TNF+ cells. Data are quantified from three independent experiments (n = 9-10 mice/group, p = 0.1188) E-F) pmel T cells from tumor-draining lymph nodes were restimulated with gp100 peptide for 1 hour and analyzed by flow cytometry for E) % p-S6+ (n = 8-10 mice/group from 3 independent experiments p = 0.8470) and F) % p-ERK+ (n = 8-10 mice/group from 3 independent experiments p = 0.7162) Statistical significance for all indicated comparisons in this figure was calculated using a two-tailed unpaired t-test. Source numerical data and unprocessed blots are available in source data.
