Immunostaining and dissection (unless said otherwise) were performed using protocols described in Jung et al., 2005 (link); Mandal et al., 2007 (link); Mondal et al., 2011 (link) using primary antibodies: mouse anti-c-Rel (1:50, a gift from N.Silverman; Stöven et al., 2000 (link)), mouse anti Relish (1:50, 21F3, DSHB), mouse anti-Antp (1:10, 8C11, DSHB), mouse anti-Wg (1:3, 4D4, DSHB), mouse anti-P1 (1:40, a gift from I. Ando), rabbit anti-Ance (1:500, a gift from A. D. Shirras; Hurst et al., 2003 (link)), rat anti-Ci (1:5, 2A1, DSHB), mouse anti-singed (1:20, Sn7C, DSHB), mouse anti-enabled (1:30, 5G2, DSHB), rabbit anti-PH3 (1:150, Cell Signaling), rabbit anti-Hh (1:500, a gift from P. Ingham; Forbes et al., 1993 ), mouse anti-Hindsight (1:5, 1G9, DSHB), mouse anti-EcR common (1:20, DDA2.7, DSHB), mouse anti-β-PS (1:3, CF.6G11, DSHB), rabbit-anti-GFP(1:100, 2555, Cell Signalling), and rat anti-shg (1:50, DCAD2, DSHB). Secondary antibodies used in this study are as follows: mouse Cy3, mouse FITC, mouse Dylight 649, rabbit Cy3, (1:500), and rabbit-FITC (1:200) (Jackson Immuno-research Laboratories).
Tissues were mounted in Vectashield (Vector Laboratories) and then followed by confocal microscopy (LSM, 780, FV10i, LSM 900).
Tissues were mounted in Vectashield (Vector Laboratories) and then followed by confocal microscopy (LSM, 780, FV10i, LSM 900).