Mouse α dpmpk 1

Primary antibodies were used at the following dilutions for immunofluorescence: chicken α-GFP (1:400; Abcam, Cambridge, MA), rabbit α-SYP-1 (1:200;(MacQueen et al., 2002 (link)), guinea pig α-HTP-3 (1:400; (Goodyer et al., 2008 (link)), rabbit α-AIR-2 (1:100;[de Carvalho et al., 2008 (link)]), rabbit α-LAB-1 (1:300;[de Carvalho et al., 2008 (link)]), mouse α-dpMPK-1 (1:500; Sigma, St. Louis, MO) and rabbit anti-RhoA (1:200; Santa Cruz). The following secondary antibodies from Jackson ImmunoResearch (Jackson ImmunoResearch, WestGrove, PA) were used at a 1:200 dilution: α-chicken FITC, α-rabbit Cy5, α-mouse FITC, and α-guinea pig FITC. Vectashield containing 1μg/μl of DAPI from Vector Laboratories was used as a mounting media and anti-fading agent.
Primary antibodies were used in the following dilutions for western blot analysis: chicken α-GFP (1:2000; Abcam), rabbit α-SYP-2 (1:200;[Colaiácovo et al., 2003 (link)]), rabbit α-SYP-3 (1:200; [Smolikov et al., 2007b (link)]), mouse α-dpMPK-1 (1:500; Sigma), mouse α-tubulin (1:2000; Sigma) and rabbit α-MPK-1 (1:2000; Santa Cruz, Dallas, TX). HRP-conjugated secondary antibodies, donkey anti-chicken, rabbit anti-mouse, and mouse anti-rabbit from Jackson ImmunoResearch were used at a 1:10,000 dilution.

L4 homozygote larvae were picked and aged to adults. The mouse α-dpMPK-1 (Sigma) was used as a primary antibody (1∶1000). Mouse α-tubulin (1∶000; DSHB) was used as a loading control. Secondary antibodies used were α-mouse antibody conjugated to horseradish peroxidase (HRP; 1∶10,000). PBST-5% milk was used for incubation and blocking. Quantification was done on FIJI [89] (link).