Figure 5
From: Numb-dependent integration of pre-TCR and p53 function in T-cell precursor development
Nuclear degradation of Numb. (a) SCB29 or SCIET27 cells were treated with CHX in a time-course study (0–2–4 h) before lysis. Whole-cell lysates were revealed in western blot with anti-numb and anti-β-actin antibodies. (b, left panel) SCB29 cells were treated with CHX at different times in the absence or presence of Rottlerin and whole cell lysates were revealed in western blot with anti-numb and anti-β-actin antibodies. (Right panel) Relative quantification of Numb modulation determined by optical densitometry (OD); quantification was normalized using β-actin levels. (c) Nuclear and cytosolic fractions of SCB29 cells treated with Rottlerin (Rott.) or DMSO, or Rottlerin plus MG132 for 12 h were revealed in western blot with anti-numb antibody, using anti-tubulin and anti-laminB antibodies as the cytosol and nucleus markers, respectively. (d, left panel) SCIET27 cells were treated with CHX at different times in the absence or presence of PMA and whole cell lysates were revealed in western blot with anti-numb and anti-β-actin antibodies. (Right panel) Relative quantification of Numb modulation determined by OD. **P-value <0.02 and *P-value <0.05. C, cytosolic fraction; N, nuclear fraction. All data are representative of three independent experiments
