Overexpression of E2F1 can induce ARF and p53 and inhibit pol III transcription

<p><b>Copyright information:</b></p><p>Taken from "RNA polymerase III transcription is repressed in response to the tumour suppressor ARF"</p><p></p><p>Nucleic Acids Research 2007;35(9):3046-3052.</p><p>Published online 16 Apr 2007</p><p>PMCID:PMC1888803.</p><p>© 2007 The Author(s)</p> F9 cells were co-transfected in duplicate with pVA1 (1 μg), pGST (1 μg) and 15 μg of empty vector (control) or vector encoding full-length E2F1, as indicated. RNA and protein were isolated 48 h after transfection. () Western blot of exogenous E2F1 using anti-E2F1 (human) antibody KH95 and of endogenous p19 using anti-p19 antibody ab80, p53 using anti-p53 antibody 1C12 and actin, to confirm equal loading of protein, using anti-actin antibody C11. () Primer extension analysis of extracted RNA using primers specific for VA1 (upper panel), and GFP (lower panel)

Induction of ARF in NARF2 cells leads to stabilization of p53 and repression of pol III transcription

<p><b>Copyright information:</b></p><p>Taken from "RNA polymerase III transcription is repressed in response to the tumour suppressor ARF"</p><p></p><p>Nucleic Acids Research 2007;35(9):3046-3052.</p><p>Published online 16 Apr 2007</p><p>PMCID:PMC1888803.</p><p>© 2007 The Author(s)</p> () NARF2 cells were treated with IPTG (1 mM) to induce ARF expression 24 h prior to harvesting. Western blot of extracted protein using anti-p14 antibody FL-132, anti-p53 antibody DO-1, anti-p21 antibody C19 and anti-actin antibody C11, as indicated. () RT-PCR analysis was performed on cDNAs generated by reverse transcription of total extracted RNA using primers specific for tRNA and ARPP P0 mRNA, as indicated. () Parental U2OS cells were treated with 1mM IPTG for 24 h prior to harvesting and RT-PCR analysis was performed using primers specific for tRNA and ARPP P0 mRNA, as indicated

Optimizing metastatic-cascade-dependent Rac1 targeting in breast cancer: Guidance using optical window intravital FRET imaging

Assessing drug response within live native tissue provides increased fidelity with regards to optimizing efficacy while minimizing off-target effects. Here, using longitudinal intravital imaging of a Rac1-Forster resonance energy transfer (FRET) biosensor mouse coupled with in vivo photoswitching to track intratumoral movement, we help guide treatment scheduling in a live breast cancer setting to impair metastatic progression. We uncover altered Rac1 activity at the center versus invasive border of tumors and demonstrate enhanced Rac1 activity of cells in close proximity to live tumor vasculature using optical window imaging. We further reveal that Rac1 inhibition can enhance tumor cell vulnerability to fluid-flow-induced shear stress and therefore improves overall anti-metastatic response to therapy during transit to secondary sites such as the lung. Collectively, this study demonstrates the utility of single-cell intravital imaging in vivo to demonstrate that Rac1 inhibition can reduce tumor progression and metastases in an autochthonous setting to improve overall survival. </p