Fig. 4: Expression of chondroitin sulfate A is controlled by AR in prostate cancer. | Nature Communications

Fig. 4: Expression of chondroitin sulfate A is controlled by AR in prostate cancer.

From: Reformation of the chondroitin sulfate glycocalyx enables progression of AR-independent prostate cancer

Fig. 4

a Schematic of CS chains, their sulfations sites C4S and C6S. b CS was purified from paraffin fixed PC patients’ samples using chondroitinase ABC enzyme (Chase) and subjected to LC-MS analysis. The C0S, C4S, and C6S mole percentages were calculated as the percentage of the total moles of disaccharides (C0S + C4S + C6S). Data are shown as the average percentage of moles of 16 PC patients. c Quantification of C4S and C6S of tissues from androgen deprivation therapy- treated (ADT, n = 8) and untreated (UT, n = 8) high-risk PC patients using LC-MS; mean ± SEM; Two tailed t test . d LC-MS C4S levels in AR (+) cell lines (LNCaP, VCaP, 22RV1) and AR (−) cell lines (PC3, Du145, IGR-CaP1). n = 9 samples (three cell lines and three biologically independent replicates); mean ± SEM; p = 0.0003 by two-tailed t test. e LC-MS C4S quantification in LNCaP cells after 72 h of AD condition and 48 h of treatment with 1, 5, or 10 nM of R1881. AD n = 6; FBS n = 5, p < 0.0001; AD + R18[1 nM] n = 9, p = 0.05; AD + R18[5 nM] n = 9, p = 0.014; AD + R18[10 nM] n = 9 biologically independent samples, p = 0.0006 by one-way ANOVA followed by Dunnett correction; data represented as mean ± SEM. f CSA expression levels in LNCaP cells in ambient conditions and AD conditions (7 days) determined by flow cytometry using rVAR2 lectin; linear regression followed by Ancova test; p = 0.0272, F = 5.529, DFn = 2, DFd = 9. g CSA and AR expression levels in LNCaP cells in ambient and AD conditions (7 days) determined by western blotting using rVAR2-V5 lectin and V5 antibody. h CHST11 mRNA levels 48 h after CHST11 knockdown. Data are shown as fold change (FC) mean ± SD; n = 3 biological replicates. i CSA expression levels in PC3 cells 48 h after siCHST11 determined by flow cytometry using rVAR2 lectin and anti-V5 antibody. linear regression followed by Ancova test; p = 0.0448, F = 6.387, DFn = 1, DFd = 6. Representative data of three independent experiments. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001. Source data are provided as a Source Data file.

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