Fig. 3

Decreased expression of NEU1 during EMT. a HCV29, KK47, and J82 cells were treated with 5 ng/mL TGFβ for 48 h. Total protein was extracted with RIPA buffer, subjected to SDS-PAGE, and transferred to PVDF membrane. Membranes were incubated with antibodies against NEU1, FN, and N-cadherin, incubated with appropriate HRP-conjugated secondary antibody, and proteins were revealed with Super-signal Chemiluminescence substrate kit as described in Supplementary information / Methods. β-tubulin: loading control. b Enzymatic activities of total sialidases in TGFβ-treated cells as described in Fig. 2A. c Expression of NEU1 at the mRNA level in TGFβ-treated and nontreated cells. d Sialic acids in TGFβ-treated and nontreated cells were stained by Cy3-labeled SNA, MAL-II (upper, middle), or by immunofluorescence staining with anti-NEU1 (lower). Scale bar: 100 μm. *, P = 0.01–0.05. **, P = 0.001–0.005. ***, P < 0.001