Anti antp

Lymph glands were dissected and processed as previously described [11 (link)]. The following primary antibodies were used: mouse anti-Col [10 (link)]; anti-proPO (1/200; T. Trenczek, Justus-Liebig-University Giessen, Giessen, Germany); anti-P1 (1/30; I. Ando, Institute of Genetics, Biological Research Center of the Hungarian Academy of Science, Szeged, Hungary); anti-Antp (1/100) and anti-Hindsight (Hnt)/pebbled (peb) (1/100) (Developmental Studies Hybridoma Bank). Rabbit anti-H3P (1/200; Upstate Biotechnology); anti-αPS4 (1/200, [11 (link)]. For in situ hybridization, DIG-labeled antisense RNA probes against col [10 (link)], tep4 [11 (link)] and latran (lat)/eye transformer (ey) [34 (link)] were used. In situ hybridized samples were mounted in 1XPBS-60% glycerol. Immunostained samples were mounted in Vectashield medium (Vector Laboratories) and analyzed with laser scanning confocal microscopy (Leica SP5). Cell nuclei were visualized using the dye TOPRO3 (Molecular Probes).

The following primary antibodies were used at the indicated dilutions for immunolabeling: Rabbit anti-β-galactosidase (Bio-Rad; 1:100), rabbit anti-Eve (GenScript; 1:1,000), chicken anti-mCherry (Novus; 1:1,000); chicken anti-GFP (Aves; 1:1,000). Mouse anti-Ubx (1:200), anti-Abd-B (1:400), anti-Antp (1:400), anti-nc82 (1:30), anti-ChaT (1:100), and anti-elav (1:100) were all obtained from the Developmental Studies Hybridoma Bank (University of Iowa).