Fig. 6: Genetic perturbation of NGFR modulates T cell sensitivity.

a Western blot analysis of indicated cell lines transduced with a control hairpin or one of two short hairpins targeting NGFR. Vinculin was used as a loading control. One blot of two independent replicates is shown (the other can be found in Source data). b Quantification of MART-1 T cell cytotoxicity in TR cell lines relative to Ctrl T cells (1:1 ratio tumor cell: T cell) in the presence of absence of shNGFR. Cells were loaded with 100nM MART-1 peptide prior to the assay, and cytotoxicity was normalized to shCtrl cell survival after MART-1 T cell attack. Error bars represent S.D. of four independent experiments with two technical replicates. Statistical analysis by Kruskal–Wallis test; *p < 0.05, ****p < 0.001. c Western blot analysis on D10 and M009R.X1.CL cells transduced with a control ORF or an NGFR-RFP ORF. GAPDH was used as a loading control. One blot of two independent replicates is shown (the other can be found in Source data). d Quantification of cell survival by flow cytometry after T cell attack in vitro in control or NGFR-ORF cells. Error bars represent S.D. of four independent experiments. Statistical analysis by Mann–Whitney, *p < 0.05. e mRNA expression of BDNF in parental and TR cells. Error bars represent S.D. of pooled analysis of three independent replicates. Analysis by unpaired t-test; *p < 0.05. f Quantification of cytotoxicity in TR cell lines relative to shCtrl cells after T cell attack (1:1 ratio tumor : T cell) in the presence or absence of shBDNF. Error bars represent S.D. of six independent experiments. Statistical analysis by Kruskal–Wallis test; *p < 0.05, **p < 0.01. Source data are provided as a Source Data file.