Fig. 2
WDFY2 localizes to endosomal tubules. a Sequential images showing GFP-WDFY2 localization to newly formed tubular structures and GFP-WDFY2 accumulation on the base of the tubules. Shown are frames from a timelapse sequence with images acquired every 2 s. Representative image from 200 observed endosomes. Scale bar: 0.5 µm. b dSTORM image showing GFP-WDFY2 localization to tubules. Representative image of five cells. Scale bar: 0.5 µm. c DNA-PAINT image showing GFP-WDFY2 and HRS localization to endosomes. Representative image of three cells. Scale bar: 0.5 µm. d Sequential images of endogenously NLAP-tagged WDFY2 showing localization to endosomes positive for mCherry-EEA1 and endosomal tubules. Shown are images from a time-lapse movie with images acquired every second. Representative image from 31 movies. Scale bar: 5 µm. e Localization of GFP-WDFY2 and mCherry-CORONIN1B on a tubulating endosome. CORONIN1B localizes to the base of WDFY2-positive tubules. The graph shows the normalized fluorescence intensity of GFP-WDFY2 and mCherry-CORONIN1B along the indicated line. Representative image from 50 observed endosomes. Scale bar 10 µm/1 µm (inset). f Localization of GFP-WDFY2 and mCherry-WASH on a tubulating endosome. WASH localizes to the base of WDFY2-positive tubules. The graph shows the normalized fluorescence intensity of GFP-WDFY2 and mCherry-WASH along the indicated line. Representative data from 50 observed endosomes. Scale bar 10 µm/1 µm (inset). g Localization of mCherry-WDFY2 and GFP-FAM21 on a tubulating endosome. FAM21 localizes to the base of WDFY2-positive tubules. The graph shows the normalized fluorescence intensity of mCherry-WDFY2 and GFP-FAM21 along the indicated line. Representative data from 50 observed endosomes. Scale bar 10 µm/1 µm (inset). h Quantification of WDFY2 tubules containing CORONIN1B, WASH, or FAM21. Results are shown as percentage of WDFY2 tubules positive for CORONIN1B, WASH or FAM21 per cell (20 cells per condition, CORONIN1B = 332 tubules, WASH = 338 tubules, FAM21 = 309 tubules). Plotted are the percentage of double-positive tubules per cell and the mean ± 95% CI. Source data are provided as a Source Data file
