Additional file 4: Figure S4.

The effect of JIP3 knockdown was offset by JIP3-WT but not JIP3-LZ4A. (A). Lysates prepared from RFP-JIP1-Neuro2a cells expressing shRNA targeting JIP3 (shJIP3), non-silencing control (NS) shRNA, TAP-JIP3-WT, or TAP-JIP3-LZ4A, in the combination as indicated were immunoprecipitated with anti-RFP antibody and analyzed by WB with the indicated antibodies. Input, cell lysate used for the immunoprecipitation assay. IP:RFP, immunoprecipitated proteins. (B). Quantification of kinesin-1 binding by RFP-JIP1 in (A). Kinesin-1 binding was normalized to the amount of precipitated RFP-JIP1 and input of kinesin-1. Results of two independent experiments are shown. (C). Differentiated RFP-JIP1-Neuro2a cells were transfected with shRNA vectors (NS or shJIP3) containing a GFP expression cassette. Arrowheads indicate the neurite tips of transfected cells. Scale bar = 20 μm. (D). Quantification of the relative fluorescence of RFP-JIP1 in the neurite tip. *: p < 0.03. Error bars indicate ± SEM. n = 50 for each construct.

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Satake et al. BMC Cell Biology 2013 14:12   doi:10.1186/1471-2121-14-12