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. Author manuscript; available in PMC: 2014 Jan 15.
Published in final edited form as: Cancer Res. 2013 Jan 15;73(2):662–671. doi: 10.1158/0008-5472.CAN-12-0653

Figure 2. Twist1 is necessary and sufficient to induce CCL2 expression.

Figure 2

A) Analysis of secreted cytokines by HMLE-Ctrl and HMLE-Twist1 cells by cytokine expression array. Conditioned media were collected for 48 hr and incubated with array membranes to measure 174 human cytokines. CCL2 levels are highlighted on one set of membranes. B) Levels of CCL2 protein were measured by ELISA in conditioned media from indicated HMLE cells. C) Levels of CCL2 mRNA were measured by QPCR analysis inindicated HMLE cells. D) Levels of CCL2 mRNA were measured by QPCR analysis in HMLE-Twist1-ER cells (solid line) or HMLE cells (dashed line) treated with 20 nM 4-hydroxy-tamoxifen. E) Lysates from 168FARN cells expressing two shRNAs against Twist1 or a control shRNA were analyzed by SDS-PAGE and probed for Twist1 and α-Tubulin (full-length blots Supplementary Fig. S2D). F) The levels of CCL2 were measured by ELISA in conditioned media from indicated 168FARN cells. G) Levels of CCL2 mRNA as measured by QPCR analysis in indicated 168FARN cells.