Skip to main content
. Author manuscript; available in PMC: 2020 Jul 1.
Published in final edited form as: Dev Cell. 2019 May 16;50(1):57–72.e6. doi: 10.1016/j.devcel.2019.04.035

Figure 3. Tubule width is a measure for glycocalyx height and can be extracted with high throughput from 2D SR reconstructions.

Figure 3.

(A) Effect of projection of a 3D tubule into 2D. Three exemplary cylinders (i)-(iii) as proxies for membrane tubules are shown. The inner diameter corresponds to the membrane-to-membrane diameter of a tubule, and the difference between total and inner diameter corresponds to the glycocalyx height on the tubule. Black dots are sampled localizations from exemplary cylinders, turquoise dots incorporate localization precision, and red lines represent the fit. (B) Analysis routine to extract tubule widths from SR reconstructions. (C) ldlD CHO cells have a mutation in the GALE gene which leads to the absence of the key sugars UDP-Gal and UDP-GalNAc. The phenotype can be rescued by supplementation of the media with GalNAc and galactose. (D) Confocal analysis of non-rescued, partly rescued, and fully rescued Sia-labeled ldlD CHO cells. Contrast settings equal between all images. (E) Sia-determined tubule width of ldlD CHO cell rescue panel. Sugar supplementation was performed at the concentrations shown for 2 days prior to labeling and analysis. Error bars: SEM; * = p < 0.05. (F) Tubule width measurements of lipid labeled ldlD CHO cells. (G) Flow cytometry of starved and rescued ldlD CHO cells showing total AF647 fluorescence from cells labeled as in Figure 3E.