P rb1 s807 811

For Western blotting, antibodies were used according to supplier guidelines. Antibodies included those specific for p-RB1-S807/811 (1:1000, no. 8516) and RB1 (no. 9309 and no. 9313, 1:500) from Cell Signaling Technology; human p16 (ab108349, 1:1000) and GAPDH (1:5000, no. Ab8245) from Abcam; mouse p16 (sc-1207, 1:200) from Santa Cruz Biotechnology; HRP-coupled secondary antibodies (nos. NA934 and NA935, 1:2000) from Sigma. RABL6A antibodies (monoclonal and polyclonal) were produced in the Quelle laboratory [13 (link),24 (link)]. For immunofluorescence, antibodies included: p-RB1-S807/811 (no. 8516, 1:1000) from Cell Signaling Technology; p16 (no. Ab108349, 1:100) from Abcam, RABL6A (8D1, 1:10) from Quelle laboratory, and secondary fluorescent-conjugated antibodies (Alexa Fluor 488 and 647, 1:5000). DAPI, to detect nuclei, was co-stained using Invitrogen ProLong Diamond Antifade Mountant with DAPI (P36962).

The primary antibodies used were anti-Cyclin D1/2 (Proteintech), anti-Cyclin E1/2 (Proteintech), anti-P21/P27 (Proteintech), anti-NKRF (Proteintech or Bethyl), anti-ZMYND8 (Proteintech), anti-ACTB (Proteintech), anti-FLAG (Proteintech), CDK2 (Proteintech), p-CDK2 Thr160 (Cell Signaling Technology), RB1 (Cell Signaling Technology), and p-RB1 S807/811 (Cell Signaling Technology). The HRP-linked anti-mouse (1/5000) and anti-rabbit antibodies (1/5000) were purchased from Cell Signaling Technology.