Rabbit anti p histone h3

Immunostaining of brain sections or dissociated cells was performed as described previously^{26 (link),35 (link),64 (link)}. Primary antibodies used were mouse anti-ARID1B (Abcam, ab57461; Abnova, H00057492-M02), rabbit anti-cleaved caspase-3 (Cell Signaling Technology, #9664), mouse anti-BrdU (BD Biosciences, #555627), rabbit anti-p-histone-H3 (Cell Signaling Technology, #9701), rabbit anti-Ki67 (Cell Signaling, #9129), rabbit anti-β-catenin (Cell Signaling, #8480), chicken anti-GFP (Invitrogen, A10262), mouse anti-Parvalbumin (Sigma-Aldrich, MAB1572), rabbit anti-Cux1 (Sigma-Aldrich, SAB2105137), and rabbit anti-Tbr2 (Sigma-Aldrich, AB2283). Appropriate secondary antibodies conjugated with Alexa Fluor dyes (Invitrogen, A32731, A32727, A32723, A32732) were used to detect primary antibodies. DAPI (Sigma-Aldrich, D9542) was used to stain nuclei. No antigen retrieval was performed in this study.

Mouse anti-Cyclin B1 (#4135), rabbit anti-CDK9 (#2316), rabbit anti-Aurora A (#3092), rabbit anti-phospho-pRb (#9307), mouse anti-pRb (#2692S), rabbit anti-MCM2 (#3619) and rabbit anti-pHistone-H3 (#9701) were purchased from Cell Signaling. Mouse anti-CDC7 (ab10535), rabbit antiphospho-MCM2 (ab70371), rabbit anti-SNRPD2 (ab155030), rabbit anti-SNRPD3 (ab111094) and rabbit anti-NHP2L1 (ab95958) were obtained from Abcam. Mouse anti-Cyclin D1 (no. sc20044) was purchased from Santa Cruz Biotechnology. Mouse anti-GFP (#11814460001) was purchased from Roche. Rat anti-phospho-RNA polymerase II (no. 04–1571) was from Merck Millipore. Mouse anti-tubulin (no. T-9026) was purchased from Sigma-Aldrich. HRP, Alexa-488 and Alexa-647 tagged secondary antibodies were purchased from Jackson Immunoresearch Laboratories.

Staining was carried out using standard protocols. Briefly, wing discs from third instar larva were dissected and fixed in PBS‐T containing 4% formaldehyde for 20 minutes at room temperature. After washing with PBS‐T, they were blocked with 5% BSA in PBS‐T for 30 minutes and then incubated with primary antibodies overnight at 4°C. Washed discs were subsequently incubated with secondary antibodies. The following primary antibodies were used: mouse anti‐MMP1 (3A6B4, 1:300) and mouse anti‐Wg (4D4, 1:500) were purchased from Developmental Studies Hybridoma Bank. Mouse anti‐β‐galactosidase (sc‐65670, 1:500) was from Santa Cruz. Rabbit anti‐Caspase3 (#9661, 1:1000) and rabbit anti‐p‐Histone H3 (#9701, 1:1000) was from Cell Signaling Technology. Secondary antibodies goat anti‐mouse Alexa Fluor594 (A11012, 1:1000) and goat anti‐rabbit Alexa Fluor594 (A11005, 1:1000) were from Invitrogen. Fluorescence images were recorded using a Nikon DS‐Ri1 fluorescence microscope and a Zeiss LSM 880 confocal microscope. Images of the adult eyes were acquired using a Nikon SMZ‐745T trinocular stereo microscope. Images were then analysed using Zeiss Zen, Image J and Adobe Photoshop software.