Extended Data Fig. 6: Analysis of the efficiency of vector delivery and T cell activities in HCC.
From: Tumor-specific cholinergic CD4+ T lymphocytes guide immunosurveillance of hepatocellular carcinoma
a,b Representative fluorescence microscopy images (a), and quantification of the proportion of hepatocytes positive for the Rosa26Confetti RFP marker (b), in liver sections from Rosa26Confetti/+;Chatfl/fl and Rosa26Confetti/+;Chatfl/fl;CD4-Cre mice on the indicated days following plasmid injection. In (b), each dot represents a microscopy image (n=6, 11, and 9 acquired on days 1, 3, and 10 from Rosa26Confetti/+;Chatfl/fl mice; n=5, 12, and 11 acquired on days 1, 3, and 10 from Rosa26Confetti/+;Chatfl/fl;CD4-Cre mice. Images were collected from 2-4 mice/group). Data are the mean ± SEM. P values were determined by two-way ANOVA with Tukey’s multiple comparisons test. c qPCR determination of transposon-derived Myc mRNA expression (relative to Actb) in livers of Chatfl/fl and CD4-Cre;Chatfl/fl mice at the indicated time points following HCC induction. The forward primer used was within the Myc ORF, with the reverse primer in the transposon vector. Each dot represents an individual mouse (n=6 Chatfl/fl mice in Control group, and n=2, 4, 3 and 5 Chatfl/fl mice on days 1, 3, 10, and 23; n=2, 4, 4, and 5 Chatfl/fl;CD4-Cre mice on days 1, 3, 10 and 23). Data are the mean ± SEM. P values were determined by two-way ANOVA with Sidak’s multiple comparisons test. d,e Percentages of CD4+ T cells (d) and CD8+ T cells (e) among mononuclear cells (MNCs) isolated from HCC-bearing livers of Chatfl/fl and CD4-Cre;Chatfl/fl mice as assessed by flow cytometry, representative of three independent experiments. Each dot represents an individual mouse (n=11 mice in Chatfl/fl and n=12 mice in Chatfl/fl;CD4-Cre). Data are the mean ± SEM. P values were determined by unpaired, two-tailed t-test. f Percentage of IFN-γ+ CD8+ T cells in HCC-bearing livers of Chatfl/fl and Chatfl/fl;CD4-Cre mice as analyzed by flow cytometry. Each dot represents an individual mouse. Each dot represents an individual mouse (n=11 mice in each group). Data are the mean ± SEM. P values were determined by unpaired, two-tailed t-test. Related to Fig. 5.
