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. 2011 Oct 18;6(10):e26135. doi: 10.1371/journal.pone.0026135

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Chan et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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(A) Effects of GGTI-298 and FTI BMS-225975 on prenylation of wild-type 3xHA-RhoA (RhoA-GG, geranylgeranylated) and 3x-HA-RhoA-F (farnesylated) mutant. Panc-1 cells stably expressing 3xHA-RhoA-GG (right panels) or 3xHA-RhoA-F (left panels) were treated DMSO, BMS-225975, or GGTI-298 for 48 h. Whole cell lysates were collected and analyzed for RhoA, Rap1, and HDJ2 processing by Western blotting. (B) Expression levels of 3xHA-RhoA-F and 3xHA-RhoA-GG in Panc-1 stable cell lines used in this experiment were analyzed by immunoblotting using antibodies against the HA tag or actin (loading control). (C and D) Panc-1 cells and Panc-1 cells stably expressing 3xHA-RhoA-GG or 3xHA-RhoA-F were treated with DMSO or P61-E7 for 48 h. Whole cell lysates were collected and analyzed for protein levels of p21^CIP/WAF1 (C, top panel), p27^Kip1 (D, top panel), or Actin (C and D, bottom panels). Data shown are representative of three independent experiments. The p21^CIP1/WAF1 (C, top panel), p27^Kip1 (D, top panel), and Actin (C and D, bottom panels) bands were quantified using ImageJ. Protein Levels of p21^CIP1/WAF1 and p27^Kip1 are represented as relative level compared to the DMSO control.