Effect of Nrp1 silencing on HS3ST3B-mediated cell proliferation and survival. Parental (pEmpty) and HS3ST3B expressing (clones C and D) cells were transfected with the siRNA targeting Nrp1 (siNrp1) or negative control siRNA (siCtrl) and then cultured for 24 h in complete medium. (A) After wash, cells were collected and cultured in medium containing 1% FCS for 24 h and 48 h. At each time, the effect of HS3ST3B on the proliferation of MDA-MB-231 cells was estimated by cell counting. Results are expressed as fold changes by comparison with the cells that have been initially added into the wells. Data are means ± S.D. from three separate experiments performed independently (** p < 0.01, *** p < 0.001, significantly different when compared with the parental cells; ##
p < 0.01, ###
p < 0.001, significantly different when compared with the siCtrl-treated cells; n.s. not significantly different). (B) Cells were seeded in six well plates (2000 per well) and cultured for nine days in the presence of 1% FCS, after which the colonies were stained with crystal violet. The left panel represents the quantification of the colonies per well. Results are expressed as fold changes by comparison with the parental cells that have been transfected with siCtrl. Data are means ± S.D. from three separate experiments performed independently (*** p < 0.001, significantly different when compared with the parental cells; ###
p < 0.001, significantly different when compared with the siCtrl-treated cells; n.s. not significantly different).