Supporting Online Material


A Septin Diffusion Barrier at the Base of the Primary Cilium Maintains Ciliary Membrane Protein Distribution
Qicong Hu, Ljiljana Milenkovic, Hua Jin, Matthew P. Scott, Maxence V. Nachury, Elias T. Spiliotis, W. James Nelson

Supporting Online Material

This supplement contains:
Materials and Methods
Figs. S1 to S5
Tables S1 to S3
References

This file is in Adobe Acrobat PDF format.

Other Supporting Online Material for this manuscript includes the following:

Movie s1
FRAP the whole cilium in IMCD3 cells stably expressing Htr6SEP. Images were captured every 3 s for 10 minutes with an exposure of 500ms.

Movie s2
FRAP of the whole cilium in IMCD3 cells transiently expressing Htr6SEP. Images were captured every 10 s for 10 minutes with an exposure of 500ms.

Movie s3
FRAP of the whole cilium in IMCD3 cells stably expressing CTSPKHD1GFP. Images were captured every 10 s for 10 minutes with an exposure of 500ms.

Movie s4
FRAP of the whole cilium in IMCD3 cells transiently expressing CTSPKHD1GFP. Images were captured every 10 s for 10 minutes with an exposure of 500ms.

Movie s5
FRAP of the whole cilium in IMCD3 cells transiently expressing Sstr3GFP. Images were captured every 10 s for 10 minutes with an exposure of 500ms.

Movie s6
FRAP of the whole cilium in MEFs stably expressing SmoYFP. Cells were treated with 100nM SAG for 24 hours before imaging. Images were captured every 10 s for 10 minutes with an exposure of 500ms.

Movie s7
FRAP of periciliary membrane Htr6SEP in IMCD3 cells transiently over-expressing Htr6SEP. Images were captured every 2 s for 3 minutes with an exposure of 500ms.

Movie s8
FRAP of the whole cilium in IMCD3 cells stably expressing IFT88YFP. Images were captured every 10 s for 9 minutes with an exposure of 500ms.

Movie s9
FRAP of part of the cilium in IMCD3 cells stably expressing Htr6SEP. Fluorescence is represented as heat-map images. Images were captured every 1 s for 20s with an exposure of 500ms.

Movie s10
FRAP of part of the cilium in IMCD3 cells stably expressing CTSPKHD1GFP. Fluorescence is represented as heat-map images. Images were captured every 1 s for 12s with an exposure of 500ms.

Movie s11
FRAP of part of the cilium in IMCD3 cells transiently expressing Sstr3GFP. Fluorescence is represented as heat-map images. Images were captured every 1 s for 20s with an exposure of 500ms.

Movie s12
FRAP of part of the cilium in MEFs stably expressing SmoYFP. Fluorescence is represented as heat-map images. Images were captured every 3 s for 30s with an exposure of 500ms.

Movie s13
FRAP of part of the cilium in IMCD3 cells stably expressing IFT88YFP. Fluorescence is represented as heat-map images. Images were captured every 2 s for 30s with an exposure of 500ms.

Movie s14
3-dimensional reconstruction (x-volume) of a primary cilium in IMCD3 cells fixed and stained for SEPT2 (green) and AcTub (red). Imaging was done by a super-resolution OMX system utilizing structured illumination (API DeltaVision).

Movie s15
3-dimensional reconstruction (y-volume) of a primary cilium in IMCD3 cells fixed and stained for SEPT2 (green) and AcTub (red). Imaging was done by a super-resolution OMX system utilizing structured illumination (API DeltaVision).

Movie s16
FRAP of the whole cilium in IMCD3 parental cells transiently expressing Htr6SEP. Images were captured every 10 s for 10 minutes with an exposure of 500ms.

Movie s17
FRAP of the whole cilium in shRNA1 IMCD3 cells transiently expressing Htr6SEP. Images were captured every 10 s for 10 minutes with an exposure of 500ms.

Movie s18
FRAP of the whole cilium in IMCD3 parental cells transiently expressing CTSPKHD1GFP. Images were captured every 10 s for 18 minutes with an exposure of 500ms.

Movie s19
FRAP of the whole cilium in shRNA1 IMCD3 cells transiently expressing CTSPKHD1GFP. Images were captured every 10 s for 18 minutes with an exposure of 500ms.

Movie s20
FRAP of the whole cilium in IMCD3 parental cells transiently expressing Sstr3GFP. Images were captured every 10 s for 10 minutes with an exposure of 500ms.

Movie s21
FRAP of the whole cilium in shRNA1 IMCD3 cells transiently expressing Sstr3GFP. Images were captured every 10 s for 10 minutes with an exposure of 500ms.

Movie s22
FRAP of the whole cilium in IMCD3 parental cells transiently expressing SmoYFP. Cells were treated with 100nM SAG for 24 hours before imaging. Images were captured every 10 s for 18 minutes with an exposure of 500ms.

Movie s23
FRAP of the whole cilium in shRNA1 IMCD3 cells transiently expressing SmoYFP. Cells were treated with 100nM SAG for 24 hours before imaging. Images were captured every 10 s for 18 minutes with an exposure of 500ms.

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