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. 2016 Apr 29;7(23):34371–34383. doi: 10.18632/oncotarget.9110

Figure 3. KIND1 is required for FRA1-promotion of cell migration.

Figure 3

(A) Immunoblotting of protein lysates isolated from FaDu cells transduced to express shCon, shFRA1, LacZ or FRA1DD. (B) KIND1 RT-PCR. Graph represents relative KIND1 mRNA levels + SD. (C) Diagram of Kind1 gene (NCBI reference # NG_016213.1). Two putative AP-1 response elements shown in capital letters were located around 200 bp from Kind1 gene transcription start site. (D) Kind1 ChIP-PCR with an anti-FRA1 antibody and primers underlined and shown in blue above. Graph represents fold-enrichment by FRA1 antibody compared to control IgG + SD. (E) Confirmation of FRA1 gene silencing by immunoblotting. (F) Effect of KIND1 gene silencing on cell migration. Images were taken at 0 h and 18 h after scratch-wounding. (G) Confirmation of KIND1 expression by immunoblotting. (H) Effect of KIND1 overexpression on cell migration. (I) Co-immunoprecipitation (IP) of c-Jun with FRA1. Protein lysates were collected from FaDu cells expressing HA-tagged FRA1DD, and then subject to IP with an antibody against HA and then immunoblotting for c-Jun and FRA1. (J) Immunoblotting of protein lysates collected from FaDu cells expressing LacZ or FRA1DD together with siCon or sic-Jun. Relative densitometry shown below each band was obtained after normalization to that of respective loading control.