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. 2016 Jul 1;11(7):e0158440. doi: 10.1371/journal.pone.0158440

Fig 4. MPT0B098 modulates JAK2/STAT3 pathway via SOCS3 accumulation.

Fig 4

(A) OEC-M1 and HSC-3 cells were treated with 0.25 μM of MPT0B098 for indicated times. The protein level of PIAS1, PIAS3, SOCS1, SOCS2, and SOCS3 were determined by Western blotting. GAPDH was used as loading control. (B) OEC-M1 and HSC-3 cells were tranfected with control vector (VC) and construct containing the SOCS3-coding region (SOCS3 cDNA #1, #2) for 48 hrs. The phosphorylated proteins (p-JAK2, p-TYK2 and p-STAT3) and total level of proteins (SOCS3, JAK2, TYK2 and STAT3) were determined by Western blotting. GAPDH was used as loading control. (C) Western blot analysis of SOCS3 levels after 48 hrs transfection of OEC-M1 and HSC-3 cells with control vector (VC) or two SOCS3 constructs (SOCS3 #1, #2). GAPDH was used as loading control. These SOCS3 overexpression transfactants were treated MPT0B098 for 24 hrs and caspases-3 activity was assessed. The data are represented as mean ± SE; **, p<0.01 versus vehicle control. (D) Western blot analysis of SOCS3 levels after 48 hrs transfection of YD-15 and DOK cells with control shRNA (NS) or two shSOCS3 constructs (shSOCS3 #1, #2). GAPDH was used as loading control. These shRNA transfactants were treated MPT0B098 for 24 hrs and caspases-3 activity was assessed. The data are represented as mean ± SE; ***, p<0.001 versus vehicle control.