Skip to main content
. Author manuscript; available in PMC: 2011 Mar 30.
Published in final edited form as: Oncogene. 2010 Aug 2;29(39):5427–5436. doi: 10.1038/onc.2010.276

Figure 1.

Figure 1

The phosphor-p68 down-regulated E-cadherin by upregulation of transcription of Snail1.

(A upper panel) Luciferase reporter of E-boxes (E-cadherin promoter) was cotransfected into p68 knockdown SW620 cells along with HA-p68s, wt or mutant (indicated). The luciferase activity was expressed as relative luciferase activity (numbers on top of bars) by comparing the luciferase activity of SW620 cells without p68 knockdown (NT) and without HA-p68s expression (define as 100). The lower panel shows the mRNA expression of E-cadherin under the same conditions. (B) Chromatin immunoprecipitations (ChIP) of the E-cadherin promoter by anti-HA antibody in SW620 cells with/without (p68/NT) p68 knockdown. The HA-p68s (WT or Y593F mutant) were exogenously expressed. The primers positions for PCRs were indicated by arrows. ChIP by mouse IgG and antibody against TFIIB, which binds to the GAPDH promoter, were used as controls. Inputs were PCR products from DNA extracts without ChIP. (C) & (D) Expression of Snail1 was examined by immunoblot of cell lysate (C) and RT-PCR of total RNAs (D) prepared from SW620 cells with/without (p68/NT) p68 knockdown. The HA-p68s (WT or Y593F mutant) was exogenously expressed. The expression and tyrosine phosphorylation of HA-p68s were examined by IB of immunoprecipitated HA-p68s (IP:HA). Total p68 level was detected by IB using monoclonal antibody p68-rgg (IB:p68). IB of histone 2A (H2A) was a loading control. RT-PCR detection of B2M mRNA in the RNA samples was a control for PCR reaction and loading.