Primary antibodies, Tie2 (R&D, #AF313, 1:100), Calpain1 (Sigma, #C5736, 1:100), FOXO3A (Cell Signaling, # 2497, 1:100), actin (
phalloidin, Invitrogen, #A12379, 1:500) NG2 (Millipore, #Ab5220, 1:100), CD31 (BD Bioscience, # 553370, 1:100) and Mac - 1 (BD Bioscience, # 550282, 1:50) were prepared
in 1 % BSA / PBS containing 0.3 % Triton - X 100 and incubated overnight at 4 °C.
Slides were stained with DAPI and Alexa Fluor488 -
Phalloidin (Invitrogen, Carlsbad, CA, USA) and mounted
in ProLong Gold (Invitrogen).
(A to D) Convergence of ventral muscles into an axochord (white arrows)
in Capitella, illustrated by
phalloidin stainings (A and C) and WMISH (red
in B and D) of selected axochord markers.