Fig. 2

FANCC and UNC5A are required for neurite outgrowth. a Representative microscopic images of SH-SY5Y cells stably transduced with shRNA against FANCC (FANCCi), UNC5A (UNC5Ai), or control noncoding scrambled shRNA (Control) vectors (upper panels) and following differentiation with retinoic acid (+RA at 10μM; lower panels). Cells were visualized at ×40 magnification. b Western blots showing the depletion of FANCC using either one shRNA (sh-C1) or a mixture of 3 (sh-C3) against FANCC (upper blot) and a mixture of 5 shRNA against UNC5A (sh-U; lower blot) in SH-SY5Y cells. c Control cells transfected with empty vectors; NT: untransfected cells. c UNC5A- and FANCC-depleted SH-SY5Y cells (UNC5Ai and FANCCi, respectively) induced to differentiate with RA (10μM) were labeled with anti-UNC5A (blue) and anti-FANCC (green) antibodies and phalloidin conjugates (red). The labeled cells were visualized by confocal fluorescence microscopy at ×100 magnification using a Nikon E800 microscope equipped with a C1 confocal system. d Estimated neurite length measured estimated Control (n = 15) FANCCi (n = 21) and UNC5Ai (n = 11) ***p > 0.0001