Figure 6.

Hypoxia potentiates CAF-dependent alteration of CTL-mediated cytotoxicity (a-c) The lysis of T1 melanoma cell line by LT12 CTL clone, pre-treated with normoxic or hypoxic CAF1-5 conditioned medium (CM) during 16 hrs, was evaluated by ⁵¹Cr release assay at different effector:target (E:T) ratios. Regular culture medium was used as control. Data (a-b) are the mean ± s.d. from two independent experiments performed in triplicate. Experiments in (a-b) were performed at the same time but separated in two different panels. (c) represents the mean ± s.d. of all T cell-mediated lysis experiments from (a-b) using the normoxic or hypoxic CAF CMs pre-treatment of the LT12 CTL clone. (d-e) Granzyme B expression by LT12 CTL clone after 16 hrs incubation with normoxic or hypoxic CAF CMs, measured by flow cytometry. Regular culture medium was used as control. A representative flow cytometry histogram using CAF2 CM treatment is shown in (d) and the percentage GzmB⁺ LT12 T cell after treatment with hypoxic or normoxic CAF1-5 CMs (mean ± s.d. of three independent experiments) in (e). N: Normoxia; H: Hypoxia. P values (a-e) were determined by unpaired two-tailed student’s t-test. (*p < .05)