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. Author manuscript; available in PMC: 2017 Sep 1.
Published in final edited form as: Cancer Res. 2016 Jun 30;76(17):4981–4993. doi: 10.1158/0008-5472.CAN-16-0232

FIGURE 5. Sdc1-coupled IGF1R sequesters and inactivates ASK1.

FIGURE 5

A. ASK1 immunoprecipitates from CAG or MM.1R cells treated for 6 hr with 10 μM SSTNIGF1R or 10 μM IGF1R inhibitor AG538 are probed for inactive (pY, pSer83 and pSer966), active (pThr838) or total ASK1; B. Sdc1 immunoprecipitates from CAG or MM.1R cells treated with 10 μM SSTNIGF1R or 10 μM AG538 are probed for the presence of total ASK1, inactive (pY) or active (pThr838) ASK1, active IGF1R (pY1131) or total IGF1R, and total Sdc1 (mAb B-A38); C. RNAi knock-down of ASK1 and IGF1R protein levels in CAG and MM.1R cells 48 hr after control (Ctrl) or target-specific siRNA transfection; D. Equal cell equivalents from control, ASK1 or IGF1R siRNA-transfected CAG and MM.1R cells (48 hr post-transfection) treated with or without 30 μM SSTNIGF1R for 8 hr are analyzed for levels of active JNK (pThr183/pTyr185) and total JNK; E. Active Caspase 3/7 is quantified in control, IGF1R or ASK1 siRNA-transfected CAG and MM.1R cells treated as in (D).