Figure 1.

BNIP3 is required to maintain clonogenic growth of melanoma cells. (a) Confirmation of effective BNIP3 KD in control and BNIP3 shRNA transduced B16-F10 cells on protein level (24 h after cell plating). A representative western blot (n=6) probed with anti-BNIP3 antibody and quantification below is shown. Actin is used as a loading control. Statistical analysis was performed as described in the materials and methods section. (b) The effect of BNIP3 silencing on spontaneous cell death determined by PI exclusion. Graph shows the percentage of dead cells (PI-positive) of control and BNIP3 KD conditions as a function of time after B16-F10 cell plating (12, 24 and 48 h) (n=3). (c) Representative images (n=3) showing clonogenic expansion of control and BNIP3 silenced B16-F10 cells. Photos were taken 10 days after seeding of the cells as a diluted single cell suspension