Extended Data Fig. 6: Analysis of KLK3 expression in T cells.
Flow cytometry sorting strategy (a) and statistics (b) of PSMA+ CD8 T cells. Relative expression of KLK3 in the indicated prostate cancer cell lines c) and EVs (d) derived from them. e, Examination of KLK3 protein (PSA, red) in T cells (CD8+, green) after co-culture with EVs derived from the indicated prostate cancer cell lines by fluorescence microscopy. Scale bars, 10μm. Data show representative results of two repeats. f, Immunoblot for small EV-enriched (CD81) and depleted proteins (GM130) in C4-2B EVs. EVs collected by differential centrifugation followed by density gradient purification. 1µg of protein was loaded for whole cell lysate (WCL). Same volume was loaded for S-EV fractions. g, rtPCR for the indicated probes in C4-2B small EVs collected by differential centrifugation and floated in different density fractions. tSNE view showing cells from the high-risk prostate cancer patient (SC001H), color coded by the tissue source (h) or cell type (i). Distribution of KLK3 expression in different cell types for samples from Batch 1 data (j) (n = 22,667, 7,495 and 629 cells for epi., immune and stroma groups, respectively), tumour tissue (k) (n = 700, 4,038 and 594 cells for epi., immune and stroma groups, respectively) and left LN (l) (n = 153, 1,954 and 2 cells for epi., immune and stroma groups, respectively) from Batch 2. Two-sided P values are calculated using Mann-Whitney U test. Y-axis of j-l represents natural logarithm scale. For box plots, center line represents the median and box limits represent upper and lower quartiles, and whiskers depicts 1.5× the IQR, extreme values outside of this range is shown as individual points. Statistical data relevant to Extended Data Fig. 6c-d, g are provided in the source data. Unprocessed Western Blots relevant to Extended Data Fig. 6f are provided in the source data.
