Figure 6. Silencing ΔNp63 mRNA expression substantially abrogates the tumorigenic potential of HTB-4 bladder cancer cells triggered by FA pathway impaired by elevated FAVL.

(A) Silencing ΔNp63 blocks the advantage in cell proliferation triggered by FAVL elevation under both hypoxia and normoxia (normal oxygen). HTB-4+EV or +FAVL stably-transfected cells with or without silenced ΔNp63 (Supplementary Figure 3) were plated at day 0 with an equal number. The total cell number was recounted on day 5 and plotted in the left panel for the relative cell growth. The representative images on day 5 were shown for cells growing under both normoxia and hypoxic stress (right panel). (B) Silencing ΔNp63 blocks the invasive advantage triggered by FAVL elevation. As performed previously [11], two sets of stably-transfected cell pairs (HTB-4+EV+Lenti-EV & HTB-4+EV+Lenti-shRNAi; HTB-4+FAVL+Lenti+EV&HTB-4+FAVL+Lenti-shRNAi) (Supplementary Figure 3) were grown in 140 μm DFO overnight and plated into transwells the following day in medium containing 140 μm DFO. Transwells were processed accordingly 2 hours after plating. Cells were counted with 10 random fields at a magnification of 400X. All counted cell numbers were used to plot the relative invasive potential (right panel). Representative images of the invasive cell density are shown in the left panel. Cells expressing elevated FAVL have a stronger invasive potential (image panel-3 compared to image panel-1). Silencing ΔNp63 can mitigate the invasive potential of cells carrying FAVL (image panel 4, compared to image panel 3), but not cells without FAVL elevation (image panel 2 compared to image panel-1; SD was made from three separate experiments).