Figure 1

Expression of VCAN in EOC monolayer cells and spheroids. (A) Expression of VCAN isoforms V0, V1, and V3 in the indicated EOC cell lines has been tested using QPCR. EEF1A1 served as a housekeeping gene control. (B) Expression of VCAN isoforms V0, V1, and V3 was detected in the indicated EOC cell lines cultured in monolayers or as spheroids. Ct values were normalized against those corresponding to the housekeeping gene control EEF1A1. Presented is an average of at least three independent experiments; bars designate the standard deviation. (C) Expression of versican V3 in individual cells released from monolayers (termed “single”) and spheroids of SKOV-3 was detected using Western blot. Expression of versican V3 was quantified using optical densitometry, normalized to the expression of a housekeeping gene β-tubulin, and statistically analysed using t-test between spheroid and monolayer groups. (D) Expression of versican in monolayers and spheroids of SKOV-3, OVCAR3, and OVCAR4 was assessed using immunofluorescence staining. Bar, 50 micron. Intensity of the versican staining in disaggregated spheroids and monolayers was measured using AxioVert software (Zeiss). Areas occupied by disaggregated spheroids were outlined to measure total intensity of the fluorescence signal, averaged, and plotted in the histogram. Student’s t-test was used to analyze differences in versican fluorescence staining in spheroids and monolayers.