Extended Data Fig. 1: Cxcl15 Regulation Upon AR signaling Stimulation and Inflammatory Stimuli in Prostate Tumor Epithelial Cells.
From: Castration-mediated IL-8 promotes myeloid infiltration and prostate cancer progression
a, Androgen responsive prostate tumor cells progressed from castration-sensitive (CS) to androgen responsive (pADT), and eventually developed castration-resistance (CR). CR was tumor size defined as ≥ 30% of nadir tumor volume. Top, tumor growth curve of MCRedAL tumors (n = 3 mice per group, repeated x2); bottom: treatment and harvest timeline. CTX: Castration. b, Upper: sorting strategy to isolate tumor epithelial cells from a based on their expression of mCherry and their CD45−CD11b−F4/80− phenotype; Lower: purity check of mCherry+ sorted cells. c, Histogram of log2 fold change comparisons (SD Log2 FC) between pADT and CS groups among all the microarray transcripts (n = 3 tumors per group). d, qRT-PCR quantification of Cxcl15 in Myc-CaP cells cultured at indicated concentrations of DHT for 8hrs and TNFα (50Units/ml) for 6hrs, cells cultured in androgen-free media for 48 hours before stimulation (n = 2 independently cultured replicates per condition, repeated x2). Expression levels normalized to mean ΔCT level in samples cultured in androgen free media without TNFα or DHT. e, qRT-PCR quantification of Cxcl15 in Myc-CaP WT cells expressing either nothing, scramble (Scr) shRNA, or an anti-AR shRNA (KD: knock-down) cultured in the presence of DHT (10 nM) for 8hrs and TNFα (50Units/ml) for 6hrs, cells cultured in androgen-free media for 24hrs before DHT stimulation (n = 2 independently cultured replicates per condition, repeated x2). Expression levels normalized to mean ΔCT level in WT samples cultured in androgen free media without TNFα or DHT. f, qRT-PCR quantification of Cxcl15 in Myc-CaP cells cultured at indicated concentrations of DHT for 8hrs and TNFα (50Units/ml) for 6hrs in the presence and absence of the AR blocker Enzalutamide (10 µM), cells cultured in androgen-free media for 48hrs before stimulation (n = 2 independently cultured replicates per condition, repeated x2). Bar plots represent means with SEM. Unpaired two-tailed (d-f) t-tests were performed, p-values ≤ 0.0001 (****); p-values ≥ 0.05 (ns).
