Fig. 7.

HA synthesis is associated with IHH secretion in HAS3-EVs. a Western blotting analysis of MV3 cells with GFP and GFP-mHAS3 D216A transfection. MV3 and MV3-GFP cells treated with or without doxycycline were also used as controls. The GFP band for HAS3 is near ~ 100 kDa while the band for only GFP is near ~ 35 kDa. b Western blotting analysis of MV3-HAS3 cells with or without 4MU, mannose and glucosamine treatments. Uninduced group was used as control. c 96 h HaCaT cell proliferation with or without treatment of EVs obtained from the described MV3 cell lines. Data represent mean ± S.E. of three independent experiments; **P value < 0.001, one-way ANOVA (Tukey’s test). In panels a and c, representative blots from 3 independent experiments are shown. d, e Melanoma tissue sections were stained to check for claspin immunoexpression, scale bar 100 µm. Data represent mean ± S.E. of 3–4 independent experiments in (a) and (b) and mean ± S.E. of 12–16 tissue sections score in (d). *P value < 0.05, **P value < 0.01, one-way ANOVA (Tukey’s test) in (a) and (b)