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. Author manuscript; available in PMC: 2019 Oct 4.
Published in final edited form as: Cell. 2018 Sep 13;175(2):514–529.e20. doi: 10.1016/j.cell.2018.08.033

Figure 3. Asters are ER proteins that localize to the PM in response to cholesterol.

Figure 3.

(A) Analysis of the cellular localization of full-length (1–699), B ∆GRAM (225–669), or B GRAM (1–171) Aster-B-GFP constructs with ER marker (Sec61β) in HeLa cells imaged by live-cell confocal microscopy. Scale bar 5 µm.

(B) Analysis of Aster-B-GFP (N-terminal, 1–738) localization and PM (CellMask PM stain) in A431 cells imaged by confocal microscopy. Cells were cultured 5% LPDS or treated with 200 µM cholesterol:cyclodextrin for 1 h. Images were taken from live cells. Scale bar 5 µm. See also Supplemental Movie 1.

(C) Live-cell imaging of GFP-Aster-B localization in A431 cells stably expressing BFP-KDEL (ER marker) and Cherry-CAAX (PM marker). Yellow arrows indicate ER tubules in close proximity to the PM containing foci of Aster-B expression. Results are representative of three independent experiments. Large images, scale bar = 10 µm; insets, scale bar = 2 µm. See also Supplemental Figure 4A–C.