Hcx pl apo cs 40 1.25 oil objective

Adherent cells: Indirect immunofluorescence on adherent cells was performed as previously described (Bellou, 2012 #68) using primary and secondary antibodies (listed in Materials and Methods Supplementary). Cell nuclei were stained using propidium iodide-PI (Sigma), samples were mounted in moviol-dabco, and images of nine fields were taken on a Leica TCS SP5 confocal microscope using HCX PL APO CS 40 × 1.25 OIL objective.
Vascular organoids/spheroids: Vascular organoids or spheroids consisting of 1,000 cells/spheroid were fixed in 3.7% paraformaldehyde for 1 h at RT, permeabilized with 0.2% Triton-X/0.9% gelatin solution for 1 h, and 0.5% Triton-X/0.9% gelatin solution for 15 min, and incubated with primary antibodies overnight at 4°C (Supplementary Table S2). Next day, the vascular organoids were washed 5x with 0.2% Triton-X and incubated with secondary antibodies for 1 h (Supplementary Table S2). After rinsing 5x with 0.2% Triton-X and incubation with Draq5 (Thermo Fisher Scientific) for 10 min, images were taken on a Leica TCS SP5 confocal microscope using HCX PL APO CS 40 × 1.25 OIL objective. At least 10 vascular organoids or spheroids were analyzed per experiment.