Fig. 2. Stat5b silencing inhibits ĚNO production. A) Confocal imaging of nitric oxide accumulation in control, Stat5b siRNA treated and untreated MIN6 cells using DAF-FM DA. After 72 hours siRNA transfection, cells were treated with 10ÁM DAF-FM DA for 30 minutes followed by treatment with PRL, incubated for further 30 minutes, washed with PBS, incubated with 5μM MitoSOX Red for 60 minutes, stained with DAPI and imaged at 358nm, 495 nm and 510 nm excitation. B) & C) Representation of the concentration of total nitric oxide end products (nitrate + nitrite) and nitrite released from Stat5b siRNA treated versus control siRNA treated MIN6 cells, respectively. P-value was calculated using Student's t-test on GraphPad Prism 5.0 software (n = 3).