Fig. 6.

Pharmacological and genetic blockade of autophagy in T_H9 cells enhance their IL-9-dependent antitumour functions in vivo. a WT mice were injected i.v. with 0.5×10⁶ B16-OVA melanoma cells and effector 2×10⁶ OT-II T_H9 cells differentiated with or without chloroquine, and mean numbers of lung tumour foci are depicted (mean+s.d., 5 mice per group, 3 independent experiments). b WT mice were injected i.v. with 0.5×10⁶ B16 melanoma cells and effector 0.1×10⁶ TRP T_H9 cells differentiated with or without chloroquine, and mean numbers of lung tumour foci are depicted (mean+s.d., 5 mice per group, 2 independent experiments). c, d Tumour growth (mean+s.e.m.) monitored three times a week, in B16-OVA or MC38 tumour-bearing control and Atg5^{fl/fl*CD4-Cre} mice that received control mouse IgG2a or anti-IL-9 neutralizing antibodies (10 mg per kg body weight) at days −1 and 0 and then twice a week for 20 days (mean+s.e.m.) e TILs of B16-OVA tumour-bearing control and Atg5^{fl/fl*CD4-Cre}mice isolated at day 20 after tumour cell injection and stimulated with ovalbumin peptides OVA_323–339 for 24 h. IL-9 expression from OVA_323–339 CD4 stimulated TILs analysed by qPCR, ELISA and FACS. f MC38 TILs were stimulated with 50 ng ml⁻¹ of PMA and 1 μg ml⁻¹ of ionomycin for 24 h. IL-9 mRNA expression was analysed by qPCR and ELISA (mean+s.d., 5 mice per group, 2 independent experiments). NS, not significant with P>0.05; *P<0.05; **P<0.01; ***P<0.001; ****P<0.0001. a, b One-way analysis of variance (ANOVA) test; c, d two-way ANOVA test; e, f unpaired Student’s t-test