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. 2007 Mar 22;26(8):2029–2040. doi: 10.1038/sj.emboj.7601659

Figure 7.

Figure 7

RGS12 is critical for NGF-mediated axonal growth in primary mouse DRG neurons. (A) N1E-115 mouse neuroblastoma cells were transfected with non-specific or mouse RGS12-directed siRNA and lysed 72 h post-transection for immunoblotting of RGS12 and actin. (B) Wild-type mouse embryo (E14) DRG neurons were transfected with non-specific or mouse RGS12 siRNA (along with YFP tracer), cultured for 2 days with NGF (50 ng/ml), replated, and stimulated with NGF for 1 day. Axons of YFP-positive cells were traced in camera lucida and quantitated using IPlab. RGS12 knockdown resulted in a significant decrease in longest axonal length (P<0.0001; Mann–Whitney test). (C) Bax-deficient mouse E14 DRG neurons were transfected as in panel B, cultured without NGF for 2 days, and stimulated with NGF (50 ng/ml) for 36 h. Axonal length of YFP-positive cells was quantitated as in panel B. RGS12 knockdown resulted in a significant decrease in longest axonal length (P<0.0001; Mann–Whitney test). (D) Photomicrographs of representative wild-type DRG neurons transfected with non-specific (left) or mouse RGS12 (right) siRNA. Cell bodies are denoted with gray arrowheads; scale bar represents 100 μm.