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. 2017 Oct 9;114(43):11482–11487. doi: 10.1073/pnas.1705536114

Fig. 4.

Fig. 4.

Role of IFN-γ in p40 mAb-mediated death of TRAMP-C2 cells. (A) Cells were treated with p40 mAb and control IgG for 6 h, followed by real-time PCR analysis for IFN-γ mRNA expression (aP < 0.01 vs. control). (B) Levels of IFN-γ were measured in supernatants by ELISA after 24 h of treatment (aP < 0.001 and bP < 0.01 vs. control). (C) Intracellular IFN-γ FACS was performed after live/dead cell exclusion. Phycoerythrin (PE) control was included to show the specificity. (D) MFI of IFN-γ was calculated. (E) Immunocytochemical analyses of IFN-γ (green) in TRAMP cells after 24 h of treatment. Results represent three independent experiments. (F) Levels of IL-10 were measured by ELISA after 24 h of treatment (aP < 0.05 vs. control). (G) TUNEL assay in control cells and cells treated with p40 mAb, IgG, p40 mAb plus different doses of IFN-γ-neutralizing Ab, p40 mAb plus IgG, and IFN-γ Ab TRAMP cells after 48 h of treatment. Under similar treatment conditions, LDH (H) and MTT (I) assays were performed. Results are mean ± SD of three different experiments (aP < 0.01 vs. control; bP < 0.01 vs. p40 mAb-treated cells).