Figure 2
From: Phosphorylated STAT5 regulates p53 expression via BRCA1/BARD1-NPM1 and MDM2
Knockdown of NPM1 enhances P-STAT5 expression but compromises T-STAT5 expression in the circumstance where P-STAT5 is absent. (a) 1.5 × 105 TF-1 cells were lentivirally transduced with shNPM1 using a titer of 1–3 TU per cell for 72 h. (b) HeLa cells (1.5 × 105 per condition) were transduced with either backbone or shNPM1 at a titer of two TU per cell for 72 h. (c) HEK 293T cells (1.5 × 105 per condition) were lentivirally transduced with shNPM1 using viral titers of 1–3 TU per cell for 72 h. NPM1 and T-STAT5 expression were quantified by performing densitometry. (d) TF-1 cells (1.5 × 105 per condition) were deprived of GM-CSF and transduced with shNPM1 at viral titer of three TU per cell for 72 h. (e) HEK 293T cells (6 × 105 per well) were plated on six-well plate on day 1 and transfected with either FLAG-NPM1 or FLAG-backbone vectors on day1 and then the transfected cells were collected and lysed on day 2 for immunoblotting assays on the protein expression of T-STAT5. For all assays, statistical analysis was performed based on three independent assays and bar graphs represent mean±S.D.
