Extended Data Figure 5: CLEM analysis of microlumen structure and FLIP/FRAP analysis of microluminal pools.
From: Luminal signalling links cell communication to tissue architecture during organogenesis
a, Overview of lexOP:secGFP; cxcr4b:nls-tdTomato embryo used for CLEM; two organs and migrating primordium were targeted for further processing. Scale bar, 200 μm. b, Re-sliced middle section of targeted organ centres, overlay of secGFP signal with corresponding EM slice (scale bar, 5 μm) and close-up view of microlumina. c, Close-up view of luminal cavity (green) distorted by kinocilium (blue). d, Traced tight junctions (red) and adherens junctions (orange) at three cross-sections of microlumen. e, Setup of FLIP experiment on Fgf3–GFP and secGFP pool highlighting repetitively bleached region (0.73 μm diameter, red circle) and regions used for total pool (green circle), background (grey box) and readout (blue circles) measurements. Plots show mean intensity of described ROIs over time. f, FRAP experiment on secGFP and Fgf3–GFP pools with a strip ROI. Mean normalized recovery curves (mean ± s.d., N = 7) and calculated half time of recovery. Arrow indicates start of bleaching.
