KLC1 is phosphorylated and KIF5 motors differentially localize to CB and distal axons. (A) Western blot of HEK 293T cells transfected with empty vector, GFP-KIF5A, GFP-KIF5B, or mCherry-KIF5C and probed with the KIF5 antibodies. Actin serves as loading control. (B) DRG sensory neurons infected with control (Ctrl) or with either shRNA against KIF5A or KIF5C and probed with the KIF5 antibodies. Actin serves as loading control. (C) IP of endogenous SFPQ from DRG sensory neuron protein lysate and blotted against endogenous KIF5A, KIF1A, KIF3C, and KIF3A. IgG serves as control IP, input represents 3% of 0.5 mg of protein lysate used for SFPQ IP, and total elute from the bead was run on the gel. (D) DRG protein lysates were treated with intestinal alkaline phosphatase (CIP) and blotted against KLC1. Actin serves as loading control. (E) Lysates of DRG neuronal cultures maintained in neurotrophins or starved were blotted for KLC1. Actin serves as loading control. (F) Staining of endogenous KIF5A, KIF5B, and KIF5C in DRG sensory neurons grown in microfluidic chambers. Scale bar 20 µm; TUJ1 (green), KIF5 (red). (G) Staining of endogenous KIF5A, KIF5B, and KIF5C in DRGs and sciatic nerve of post-natal day 1 (P1) mice; n = 4 independent staining of tissues; scale bar: 30 µm and 9 µm for unmagnified images and merge magnified images, respectively; Tuj1 (green), KIF5 (magenta).