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. Author manuscript; available in PMC: 2008 Apr 30.
Published in final edited form as: Cancer Cell. 2007 May;11(5):447–460. doi: 10.1016/j.ccr.2007.03.009

Figure 4. c-Jun regulates RACK1 transcription.

Figure 4

A. Go6976 inhibits JNK activation. A375 cells were treated with Go6976 for 2h before protein extracts were prepared. Levels of P-JNK and total JNK were assessed by Western blot. β-actin was used to monitor equal loading.

B. Inhibition of PKC reduces RACK1-driven luciferase activity. The luciferase activity driven by the RACK1 promoter (2Kb of the proximal RACK1 promoter cloned into a pGL2 vector) was assessed in Lu1205 cells in the presence of Go6976 (3μM for 8h). TRE and TOP vectors were used as positive and negative controls, respectively. Results are shown as the mean ± SD. Data were standardized on the basis of β-galactosidase activity.

C. c-Jun regulates RACK1 expression. Protein extracts from SW1 cells stably transfected with FLAG-TAM67 were blotted with RACK1 and FLAG antibodies. β-actin antibody was used to monitor equal protein loading.

D. RACK1 expression is reduced in c-Jun-/- fibroblasts. Protein extracts from c-Jun -/- and control fibroblasts were blotted with RACK1 antibody. β-actin and α-Tubulin antibodies were used to monitor equal protein loading.

E. Decrease in relative mRNAs levels of RACK1 in c-Jun -/- MEF and in SW1-TAM67 cells. Relative levels of RACK1 mRNA were determined by Real-Time quantitative PCR. Reactions were run in triplicate. β-actin was used as a control. Results are shown as the mean (bar) ± SD of the respective relative concentrations. A representative experiment (of three performed) is shown.

F. c-Jun regulates RACK1-driven transcription. The luciferase activity driven by the RACK1 promoter (2Kb of the proximal RACK1 promoter cloned into a pGL2 vector) was assessed in SW1 cells expressing TAM67 as well as in wt and c-Jun mutant fibroblasts. Results are shown as the mean ± SD. Data were standardized on the basis of β-gal activity.

G. Activation of the JNK/c-Jun pathway increases RACK1 expression. HEK293T cells were transfected with ΔMEKK1 or empty pEF plasmid. Protein extracts were obtained 48h post-transfection and blotted with the indicated antibodies. β-actin antibody was used to monitor loading. Arrow indicates position of ΔMEKK1.

H. ΔMEKK1 increases RACK1-driven transcription. The luciferase activity driven by the RACK1 promoter (2Kb of the proximal RACK1 promoter cloned into a pGL2 vector) was assessed in HEK293T cells transfected with ΔMEKK1. Results are shown as the mean ± SD. Data were standardized on the basis of β-gal activity.