Figure 6
From: Non-histone nuclear factor HMGB1 is phosphorylated and secreted in colon cancers
Extracellular HMGB1 promote cell invasion. (a) Immunoblot analysis of HMGB1 from the culture medium of HCT116 cells transfected with NLS-WT, NLS(1,2)-AT, or NLS(1,2)-ET vectors. Secreted HMGB1 was evident in the cell lines transfected with NLS-WT and NLS(1,2)-ET. (b) Immunoprecipitation of media protein lysates using HMGB1 antibody and immunoblot using antibodies against phosphorylated proteins. Similar amount of endogenous phosphorylated HMGB1 was found in all of four cell lines transfected with different vectors. (c) Invasion activity are increased in HCT116 cells transfected with NLS(1,2)-ET vector than in cells transfected with other forms of vectors (NLS-WT, NLS(1,2)-AT and EGFPN1). Especially, the invasiveness was increased in cells transfected with NLS(1)-ET, NLS-35ET, NLS-39ET, and NLS-42ET as compared with cells transfected with the other three vectors (NLS-46ET, NLS-53ET, and NLS(1)-AT). (d) Treatment of recombinant HMGB1 protein in the culture media showed increased invasiveness of tumor cells.
