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. Author manuscript; available in PMC: 2021 Jan 1.
Published in final edited form as: Cancer Res. 2019 Oct 31;80(1):102–115. doi: 10.1158/0008-5472.CAN-19-1957

Figure 4.

Figure 4.

Inhibition of VCP enhances NIS function. A, RAI uptake of MDA-MB-231 (NIS+) and TPC-1 (NIS+) cells treated with ES-1 for 24 hours. B, Same as A, but cells treated with NMS-873. C, Western blot analysis following ES-1 or NMS-873 treatment in TPC-1 (NIS+) cells. D, PLA showing specific interaction (red fluorescent spots) between NIS-MYC and VCP in TPC-1 cells treated with ES-1. Blue, DAPI nuclear stain. Magnification, ×100. Scale bars, 10 μm. E, Co-IP assays demonstrating interaction of VCP and NIS in TPC-1 (NIS+) cells treated with ES-1 or NMS-873. F and G, RAI uptake and relative NIS protein levels in parental TPC-1 cells treated with ES-1, NMS-873, or DMSO. H, Time course of RAI uptake (top) and relative NIS protein levels (bottom) in TPC-1 (NIS+) cells treated with 2.5 μmol/L ES-1. I, Same as H, but cells treated with 5 μmol/L NMS-873. J, RAI uptake and Western blot analysis in TPC-1 (NIS+) cells transfected with VCP or Scr siRNA, then treated with ES-1. K, RAI uptake and Western blot analysis in MDA-MB-231 (NIS+) cells, TPC-1 (NIS+) cells, and human primary thyrocytes transfected with WT VCP or QQ VCP mutant. NS, not significant; *, P < 0.05; **, P < 0.01; ***, P < 0.001.