Mouse anti armadillo n2 7a1

All procedures (TEM and Immunofluoresence) were performed as previously described [14 (link), 15 (link)]. The primary antibodies used in this study were: mouse anti-elav (9F8A9, 1:200, Developmental Studies Hybridoma Bank); mouse anti-Crumbs (Cq4, 1:100, Developmental Studies Hybridoma Bank), rabbit anti-aPKC ζ (zeta) (1:200, SAB4502380 Sigma), mouse anti-HA (6E2, 1:500, Cell Signaling), mouse anti-myc (9B11, 1:500 Cell Signaling), rat anti-Crumbs (1:500) [73 (link)], mouse anti-armadillo (N2 7A1, 1:100, Developmental Studies Hybridoma Bank). The C-terminal peptide KVNKLISRFEGGRPRLC (produced in the laboratory of Dr. Charles Zuker) was used as the antigen in rabbits, and the resulting antibody was used at a concentration of 1:200. Fluorescent conjugated secondary antibodies were obtained from either Jackson ImmunoResearch Laboratories or Life Technologies. Rhodamine or Alexa Fluor 647 conjugated phalloidin (1:200, Life Technologies) were utilized for the detection of F-Actin. Confocal images were captured on a Leica TCS SP5. TEM imaging was conducted with a JOEL 1010 and JOEL 1400. All images were processed in Adobe Photoshop.

Eye-antennal imaginal discs and salivary glands were prepared for immunohistochemistry using standard protocols. As the growth conditions strongly affect salivary gland size, all the experiments were controlled by synchronization of L3 wandering larvae after a single-day egg collection. To further control this issue, a controlled the egg laying for 5 h was set up, and the salivary glands were analysed 96 h after egg laying (96–101 h AEL; electronic supplementary material, figure S4g–j′).
Primary antibodies used were: mouse anti-Armadillo N27A1 at 1 : 100 (Developmental Studies Hybridoma Bank, DSHB), mouse anti-Dlg at 1 : 1000 (4F3, DSHB), rabbit anti-Viriato (Vito) at 1 : 250 (ABGent), rat anti-DCad at 1 : 100, mouse anti-AH6 at 1 : 10 (DSHB), rabbit anti-Fibrillarin at 1 : 250 (Abcam, #ab5821), mouse anti-Fibrillarin at 1 : 500 (Abcam, #ab4566), mouse anti-RpS6 at 1 : 100 (Cell Signaling, #2317), mouse anti-RpL11 at 1 : 100 (Abcam, #ab79352), mouse anti-RpL10A at 1 : 400 (Abcam, #ab55544), rabbit anti-RpL22 at 1 : 100 (kind gift from Dr Vassie Ware). To stain for cellular limits phalloidin conjugated with rhodamine was used at a dilution of 1 : 1000. Appropriate Alexa-Fluor conjugated secondary antibodies were from Molecular Probes. Images were obtained with the Leica SP2 confocal system or Leica SP5 confocal system and processed with Adobe Photoshop.

Ovary dissection, fixation and staining were performed using standard protocol [58 (link)]. The following primary antibodies were used: mouse anti-Armadillo [N27A1] [Developmental Studies Hybridoma Bank (DSHB), 1:50; rat anti-DE-Cadherin (DCAD2) 1:50 (DSHB); rabbit anti-Zpg 1:2000 and guinea pig anti-Inx2 1:1000 (gift from Guy Tanentzapf), rabbit anti-phospho-Myosin Light Chain 2 (Ser19) Antibody 1:10 (catalog No: 3671, Cell signaling), and rabbit anti- Zipper 1:200 (gift from Eric Wieschaus), mouse anti-β-Gal (40-1a, DSHB) 1:100, Hindsight (1G9, DSHB) 1:5. Secondary antibodies conjugated with Alexa-488 and Alexa-568 (Molecular Probes) were used at 1:250 and 1:400 dilutions respectively. Rhodamine Phalloidin staining was performed using standard procedure [58 (link)].