Fig. 1: Altered phenotype and attenuated cytotoxicity of NK cells in the prostate tumor microenvironment.
From: C/EBPβ-dependent autophagy inhibition hinders NK cell function in cancer
A Breeding scheme for the establishment of the Pten knockout (Ptenpc−/−) mouse model. B Hematoxylin and eosin staining of prostate tissues from Ptenpc+/+ and Ptenpc−/− mice. Original magnification, ×20. Scale bar: 100 μm. Representative of an experiment of n = 4 mice/group. C Bubble plot showing the average frequency, referred to CD45+ live cell gate, of immune cell subsets in Ptenpc+/+ and Ptenpc−/− prostate (n = 4 mice/group). D Flow cytometry analysis of NK cell maturation in Ptenpc+/+ and Ptenpc−/− prostate (n = 10 mice/group). E Flow cytometry analysis of effector molecule expression in NK cells in Ptenpc+/+ and Ptenpc−/− prostate, according to gating strategy in S1G (n = 10 Ptenpc+/+ mice/group, n = 6 Ptenpc−/− mice/group). F Cytotoxicity of sorted splenic NK cells from Ptenpc+/+ mice and tumor-infiltrating NK cells from Ptenpc−/− mice (three different mice/conditions). G Tumor volume. n = 5 mice/group. H Hematoxylin and eosin staining of prostate tissues from αNK1.1 or isotype-treated mice. Original magnification, ×20. Scale bar: 100 μm. I Bubble plot showing the average frequency, referred to CD45+ live cell gate, of immune cell subsets in Ptenpc−/− prostate of αNK1.1 or isotype-treated mice (n = 5 mice/group). J Experimental scheme for tumor-conditioned NK cell generation. K Flow cytometry analysis of NK cell maturation in tumor-conditioned and control NK cells (n = 5 biological replicates, independently collected batches of tumor-conditioned medium). L Flow cytometry analysis of effector molecule expression in tumor-conditioned and control NK cells (n = 8 biological replicates, data pooled from two independent experiments). M Quantification of IFNγ release (n = 4 biological replicates/group). N Experimental scheme for cytotoxicity assay. O, P Cytotoxicity of tumor-conditioned and control NK cells against YAC-1 (O) and Pten−/− (P) target cells (n = 6 biological replicates, independently collected batches of tumor-conditioned medium). Data in C and I are presented as bubble plot showing average values; two-tailed unpaired t test. Data in (D), (E), (G), (K), and (L) are presented as Min to Max box-and-whisker plot, the box extends from the 25th to 75th percentiles and the whiskers reach the sample maximum and minimum values, the median is indicated at center line and the mean value is indicated as “+”; two-tailed unpaired t test. Data in (F) are presented as scatter plot mean ± SEM; two-tailed unpaired t test. Data in (M) are presented as scatter plot with bar with mean ± SEM; two-tailed unpaired t test. Symbols in (O) and (P) represent mean and error bars indicate SEM; two-tailed unpaired t test for corresponding E:T ratio.
