Figure 5
Detection of TG2 protein under non-denaturing conditions. (A) Representative Western blots for TG2 in total cellular proteins isolated from resveratrol-treated SHYTG2, SHYmutant and Panc-28 cells present near and away from the scratch. Equal amount of proteins were separated on non-denaturing conditions and Western blots were done with anti-TG2 antibody. In addition to the major TG2 band, another TG2 protein band with lower migration was detected in resveratrol-treated SHYTG2, and Panc-28 cells present near the scratch. (B) Representative Western blots for TG2 with cytoplasmic and membrane protein fractions prepared from DMSO- or resveratrol-treated SHYTG2, and Panc-28 cells present near the scratch. Under non-denaturing conditions, TG2 protein band with slower migration was present in both cytoplasmic and membrane protein fractions prepared from resveratrol-treated (1 and 10 μM) cells. (C and D) Western blots using anti-tyrosine antibody to detect phosphorylation state of TG2 in total cell extract (C) and cytoplasmic and membrane protein fractions (D). Native proteins (100 μg) isolated from resveratrol or DMSO treated SHYTG2 and Panc-28 cells present near the scratch, separated on non-denaturing gels followed by Western blot with anti-phosphotyrosine (p-tyrosine). The bottom panel (D) represents the Western blot for TG2 in SHYTG2 cells.