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. 2022 Aug 13;13:4760. doi: 10.1038/s41467-022-32530-7

Fig. 6. Chondroitin sulfate enables cell motility and metastases in castration-resistant prostate cancer.

Fig. 6

a CSA measurements in LNCaP cells after 72 h of Fluorosamine (FL) treatment using rVAR2 binding on flow cytometry; linear regression followed by Ancova test; p < 0.0001, F = 18.59, DFn = 3, DFd = 12. b LC-MS quantification of C4S in LNCaP cells in AD condition after 72 h of FL treatment (50 μM). Data are shown as mean ± SEM; n = 4 biological replicates; p = 0.0041 by two-tailed t test. c FL EC50 values in AR + and AR- PC cell lines. Data are shown as EC50 plus upper and lower limits; n = 8 biological replicates. d Growth (IncuCyte) of LNCaP cells in FBS or AD conditions ± FL treatments. Data are shown as mean ± SEM, n = 4 biologically independent cell cultures; after 100 h p < 0.0001 by two-way ANOVA followed by Tukey’s test. e Cell death measurement of LNCaP cells treated as in d using propidium iodide (red staining; Incucyte) (Left panel). Data are shown as mean ± SEM, n = 4 independent samples; after 100 h p < 0.05 by two-way ANOVA followed by Tukey’s test. Representative image of Incucyte at endpoint (Right panel) f Representative images of wound healing closure in PC3 exposed to FL treatments, yellow show original scratch wound limits. Blue shows closed wound margins. g Scratch width measurements of f in μm. Data are shown as mean ± SD, n = 9 biologically independent replicates; after 6 h p < 0.001 by two-way ANOVA followed by Tukey’s test. h Invasion capacity (IncuCyte; Chemotaxis assay) of PC3 in presence and absence of FL treatment. Results are expressed as phase object count normalized to initial top value and shown as mean ± SD, n = 3 biologically independent replicates; after 26 h p < 0.05, after 46 h p < 0.0001 by two-way ANOVA followed by Tukey’s test. i Protocol schematic of PC3 metastatic seeding capacity in mice. j Representative H&E and HOXB13 staining of PC3 bone and lung metastases. Scale bar represents 100 μm k IVIS imaging of mice for metastases detection at humane or experimental endpoint. D is the day of termination. l Number of mice with jaw/bone or lung metastases in PBS or FL treated group. m Kaplan–Meier estimates of metastasis-free survival of mice injected with PC3 cells pretreated with FL 25 μM or Mock; Log-rank test. ns: not significant; **p < 0.01; ****p < 0.0001. Source data are provided as a Source Data file.