Sc 101526

The mouse monoclonal antibody (mAb) against mRFP (sc-101526) was from Santa Cruz Biotechnology, the mouse mAb against GM130 was from BD Transduction Laboratories (610822), and the mAb against α-tubulin was from Sigma-Aldrich (T6199). The rat mAb against HA (11867423001) was from Roche. The rabbit polyclonal antibody (pAb) against GFP (GTX113617) was from GeneTex. Peroxidase-conjugated secondary Abs for Western blots (115-035-146, 111-035-144, and 112-035-143) and the Alexa Fluor 594-labeled secondary Ab for immunostaining (115-585-146) were from Jackson ImmunoResearch.

For whole-mount antibody staining, larvae were removed the skin with the Tweezers, washed several times with PT (PBS + 1% Triton X-100) and incubated with primary antibodies below: Anxa4 (1:1000; ab71286, Abcam, Cambridge, MA), 5-methyl-cytosine (1:100; ab10805, Abcam, Cambridge, MA), Alcam (1:50; zn5, ZIRC, Eugene, OR), Bhmt (1:500, a kind gift from Jinrong Peng, Zhejiang University, China), Dendra2 (1:1000; AB821, Evrogen, Moscow, Russia), pS6 (1:500; 2215, Cell Signaling, MA, USA), p4EBP1 (1:500; 2855, Cell Signaling, MA, USA), Dnmt1 (1:200, sc-20701, Santa Cruz Biotechnology, Santa Cruz, CA, a kind gift from Jingwei Xiong, Peking University, China), GFP (1:1000, ab6658, Abcam, Cambridge, MA), DsRed (1:500, sc-101526, Santa Cruz Biotechnology, Santa Cruz, CA) and Tomato (1:1000; orb182397, Biorbyt, TX, USA), After primary antibody incubation, larvae were washed several times with PT and incubated with secondary antibodies conjugated to Alexa Fluor 488/568/633 (1:1000; Invitrogen, Grand Island, NY). The primary and secondary antibodies were diluted in the blocking solution (PBS + 4% BSA + 1% Triton X-100) and incubated at 4 °C overnight and washed with PT for five times.