Mms 435p

The following antibodies were used. βIII Tubulin (Covance Cat# MMS-435P, RRID:AB_2313773), P53 (phospho S15) (Cell Signaling Technology Cat# 9284, RRID:AB_331464), P53 (1C12) (Cell Signaling Technology Cat# 2524, RRID:AB_331743), P53 (CM5) (Leica Biosystems Cat# P53-CM5P, RRID:AB_2744683), Puma (Cell Signaling Technology Cat# 7467, RRID:AB_10829605), Caspase-3 (8G10) (Cell Signaling Technology Cat# 9665, RRID:AB_2069872), mCherry (1C51) (Abcam Cat# ab125096, RRID:AB_11133266).
The following chemicals and biochemical reagents were used: Pifithrin-μ (Selleck Chemicals, Catalog No.S2930), ABT-737 (Selleck Chemicals, Catalog No.S1002), Caspase-Glo 3/7 Assay System (Promega, G8090), Nerve Growth Factor 2.5S (Promega, G5141).

Embryos younger than E15.5 were fixed with 4% paraformaldehyde in PBS overnight at 4°C. Embryos at E15.5 or older were intracardially perfused with 4% paraformaldehyde and post-fixed overnight at 4°C. Samples were embedded in OCT and sectioned at 16-μm thickness. Immunostainings were performed according to standard protocols. Primary antibodies were as follows: Blbp (Millipore, ABN14; rabbit, 1:500), BrdU (Abcam, ab6326; rat, 1:100), Isl1/2 (Developmental Studies Hybridoma Bank, Clone # 39.4D5; mouse, 1:500), Ki67 (Vector Laboratories, VP-RM04; rabbit, 1:250), Nf2 (Sigma, HPA003097; rabbit, 1:1000 and amplification with Invitrogen TSA kit), p75 (Promega, G3231; rabbit, 1:2000), S100β (Sigma, S2532; mouse, 1:100 and amplification with Invitrogen TSA kit), Sox10 (Santa Cruz, sc-17342; goat, 1:250), TrkA (provided by Louis Reichardt, University of California, San Francisco, CA; rabbit, 1:1000), Tuj1 (Covance, MMS-435P; mouse, 1:1000), Yap/Taz (Cell Signaling, 8418; rabbit, 1:200 and amplification with Invitrogen TSA kit), and pYap (Cell Signaling, 4911; rabbit, 1:200). Secondary antibodies were from Invitrogen and Jackson ImmunoResearch. Fluorescent images were acquired with a Zeiss LSM 510 or 780 confocal microscope.