Immunostaining was carried out as previously described [6 (link)].
Anitbodies and dilutions used were Anti-Nubbin (1:500) (gift of S. Cohen) [47 (link)], mouse anti-βgal (1:100) (DSHB; 40-1a-s), rabbit anti-βgal (1:500) (MP Biomedicals), mouse anti-GFP (1:10)(DSHB 12E6), rabbit anti-Myc (1:500) (Santa Cruz Biotech d1-717 sc-28207), rabbit anti-PH3 (1:500) (Millipore), mouse anti-Nimrod (1:1000)(gift from I. Ando)[116 (link)] and anti-Twist (1:200) (gift from A. Stathopoulos)[144 (link)].
Alexa Fluor (AF) secondary antibodies from Molecular Probes were AF488, AF555 and AF633 (used at 1:500). Nuclei were labeled with DAPI (Sigma)(1:5000).
EdU incorporation was carried out using the click-it EdU Alexa Fluor 594 Imaging kit (Molecular Probes) as previously described [145 ]. Samples were mounted in Vectashield (Vector Labs).
Immunostained samples were imaged on a Zeiss LSM 700 confocal microscope and images were processed using ZenLite, Adobe Photoshop, and Image J software. Bright-field imaging of adult wings was done on an Olympus SZX10 microscope using the CellSens Dimension software, and images were processed using Image J.
Anitbodies and dilutions used were Anti-Nubbin (1:500) (gift of S. Cohen) [47 (link)], mouse anti-βgal (1:100) (DSHB; 40-1a-s), rabbit anti-βgal (1:500) (MP Biomedicals), mouse anti-GFP (1:10)(DSHB 12E6), rabbit anti-Myc (1:500) (Santa Cruz Biotech d1-717 sc-28207), rabbit anti-PH3 (1:500) (Millipore), mouse anti-Nimrod (1:1000)(gift from I. Ando)[116 (link)] and anti-Twist (1:200) (gift from A. Stathopoulos)[144 (link)].
Alexa Fluor (AF) secondary antibodies from Molecular Probes were AF488, AF555 and AF633 (used at 1:500). Nuclei were labeled with DAPI (Sigma)(1:5000).
EdU incorporation was carried out using the click-it EdU Alexa Fluor 594 Imaging kit (Molecular Probes) as previously described [145 ]. Samples were mounted in Vectashield (Vector Labs).
Immunostained samples were imaged on a Zeiss LSM 700 confocal microscope and images were processed using ZenLite, Adobe Photoshop, and Image J software. Bright-field imaging of adult wings was done on an Olympus SZX10 microscope using the CellSens Dimension software, and images were processed using Image J.
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