mouse anti unc18 rop 4f8  (Developmental Studies Hybridoma Bank)

Journal: Journal of Cell Science

Article Title: Regulation of dendrite growth and maintenance by exocytosis

doi: 10.1242/jcs.174771

Figure Lengend Snippet: Rop interacts with the exocyst complex. (A–C) Dendritic morphology of a wild-type (A), Sec5E10 (B) or Sec6Ex15 (C) C4da-ddaC MARCM clone in a fixed preparation of late-stage third instar larva. (D,E) Quantification of the total number of terminal dendrite branches (D) and total dendrite length (E) in wild-type, Sec5E10 and Sec6Ex15 C4da-ddaC MARCM clones. ***P<0.001 (one-way ANOVA with Tukey's HSD post-hoc analysis). (F) Sholl analysis of wild-type, Sec5E10 and Sec6Ex15 C4da-ddaC MARCM clones. FRT40A and FRT42D control clones were pooled as wild type in (D–F) for the purposes of statistical analysis. (G–L) Dendritic morphology of a C4da-ddaC neuron in a wild-type (G), RopG27/+ heterozygous (H), Sec5E13/+ heterozygous (I), Sec6Ex15/+ heterozygous (J), Sec5E13/+; RopG27/+ trans-heterozygous (K) and Sec6Ex15/+; RopG27/+ trans-heterozygous third instar larva at 144 h AEL (L). All neurons were imaged live. (M) Quantification of the total number of terminal dendritic branches per 10.1×104 μm2 field. #Statistical comparison to wild type; *P<0.05, ###P<0.001 (one-way ANOVA with Tukey's HSD post-hoc analysis). (N) Rop forms a complex with exocyst components Sec6 and Sec15. Co-immunoprecipitation using Rop monoclonal antibody 4F8 precipitates Sec6 and Sec15 from adult whole-brain lysates, as detected by immunoblotting. Arrow indicates Sec6 protein (top) and Sec15 protein (bottom). Means±s.d. are shown. D,E,M, the number of neurons analyzed for each genotype are indicated.

Article Snippet: Immunocytochemistry Embryos and larvae were processed for immunocytochemistry, as previously described ( Kim et al., 2006 ), using the following antibodies: mouse anti-Rop 4F8 at 1:200 (Developmental Studies Hybridoma Bank) ( Harrison et al., 1994 ), rabbit anti-Myc at 1:1000 (A-14, Santa Cruz Biotechnology), rabbit anti-β-galactosidase at 1:5000 (Cortex Biochem), rat anti-mCD8 at 1:200 (Life Technologies), rabbit anti-Dcp-1 at 1:500 (Cell Signaling Technologies), Cy5-conjugated anti-horseradish-peroxidase at 1:250 (Jackson Laboratories), and appropriate fluorophore-conjugated secondary antibodies at 1:200 (Jackson Laboratories).

Techniques: Clone Assay, Control, Comparison, Immunoprecipitation, Western Blot