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. 2022 Sep 28;221(11):e202110132. doi: 10.1083/jcb.202110132

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© 2022 Jani et al.

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Figure S1. — PI4P and KIF13 do not associate with early sorting endosomes. (A and B) Live imaging frame of Hela cells co-expressing (A) mCh-KIF13B (magenta) and iRFP-RAB5 (pseudocolored in green), or (B) GFP-coupled PIxP sensors (green) for either PI4P (SidC-GFP, top) or PI3P (GFP-FYVE, bottom) with iRFP-RAB5 (pseudocolored in magenta). Magnified insets (4×) show RAB5⁺ structures overlapped with PI3P (B, bottom; arrowheads), but not with KIF13B⁺ tubules (A; arrows) or PI4P (B, top; arrows). Right panel shows the quantification of the shortest average distance (in µm) between mCherry-KIF13B and iRFP-RAB5⁺ structures. Data are the average of three independent experiments (>30 iRFP-RAB5⁺ structures) presented as mean ± SEM. (C) IFM of fixed Hela cells expressing the PI4P sensor (GFP-SidC, green) and labeled for markers (magenta) of TGN (TGN46; top) or plasma membrane (fluorescent-conjugated Wheat Germ Agglutinin, WGA; bottom). Arrowheads point areas of overlap. Cell periphery is delimited by yellow lines. Scale bars: (main panels) 10 µm; (insets) 2 µm.