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. 2014 Aug 10;21(5):682–699. doi: 10.1089/ars.2013.5212

FIG. 4.

FIG. 4.

Effect of α-mangostin on the expression of pStat3Tyr705, pStat3Ser725, and Stat3 DNA binding in PC cells. (A) Effect of α-mangostin on protein level of pStat3Ser727 and pStat3Tyr705 in PANC1 cells (Ai) as determined by the western blot analysis. Bar graph (Aii) represents the quantitative analysis of the western blots. (B) Effect of α-mangostin on Stat3 DNA binding activity in ASPC1 (Bi, ii) and PANC1 (Biii, iv) cells as determined by EMSA. Lane 1 NC in the blots (Bi, Bii) represents the mutant consensus sequence of Stat3. Lane 2 in blot (Bi, ii) denotes positive control (PC). Bar graphs (Bii, iv) are the quantitative analysis of EMSA blots of (Bi, ii). (C) Effect of α-mangostin on NF-κB/p65-induced constitutive activation of Stat3 in PC cells. In brief, 80% confluent ASPC1 cells were transiently transfected with NF-κB/p65 overexpressing plasmids. In a parallel set, cells were also transfected with empty vector. After 48 h transfection, cells were treated with vehicle or α-mangostin for 12 h. Whole cell lysates were prepared and western blots were performed to examine the expression of pNF-κB/p65 (Ci), total NF-κB (Ci), pStat3Ser725 (Cii), and pStat3Tyr705 (Cii). PC in the blots denotes to positive control. All blots were quantified and normalized by actin antibody. AN denotes arbitrary number, which represents quantitation of the western blots. Stat3, signal transducer and activator 3.