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. Author manuscript; available in PMC: 2018 Feb 27.
Published in final edited form as: Sci Signal. 2017 Dec 12;10(509):eaan6282. doi: 10.1126/scisignal.aan6282

Figure 6. Twist transcriptionally regulates CRP-induced cytokine expression in myeloma cells.

Figure 6

(A) Heat map of the expression of transcriptional factors in myeloma cell lines treated with or without CRP (5 μg/mL) for 24 hours. (B) ChIP assay for the interaction of Twist with the promoters of MCP-1 or RANKL using anti-Twist antibody or rabbit IgG. The input proteins served as controls. (C) Western blot for Twist abundance (top) and real-time PCR for MCP-1 and RANKL mRNAs in Twist-knockdown (KD) myeloma cells and non-targeted shRNA control (Ctrl) cells, cultured with or without CRP. (D and E) Luciferase activity in cultured ARP-1 (D) or MM.1S (E) cells transfected with a luciferase reporter-containing wild-type, truncated mutants, or Twist binding site-mutant RANKL or MCP-1 gene promoters. (F) Real-time PCR for gene expression in OC precursors cocultured with Twist-knockdown or control ARP-1 cells cultured with or without CRP. *P < 0.05, ** P < 0.01 by student’s t test. Data from four independent experiments are shown.