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. 2017 May 30;15(6):154. doi: 10.3390/md15060154

Figure 6.

Figure 6

Inhibition of pRB phosphorylation and increase of pRB/E2F complexes by fucoidan in HCT116 cells. p53+/+ (A,C) and p53−/− (B,D) HCT116 cells were treated with 150 μg/mL of fucoidan for the indicated times. (A,B) The cells were lysed, and then equal amounts of proteins were separated on SDS–polyacrylamide gels and transferred onto membranes. Membranes were probed with the indicated antibodies, and the proteins were visualized by an enhanced chemiluminescence (ECL) detection system. Actin was used as an internal control. (C,D) The cells were incubated with or without 150 μg/mL of fucoidan for 24 h, and then equal amounts of proteins (500 μg) were immunoprecipitated with the anti-E2F-1 or anti-E2F-4 antibodies. Immunocomplexes were separated by 8% SDS–polyacrylamide gel electrophoresis (PAGE), transferred to membranes, and probed with the anti-pRB antibodies.