Journal: The Journal of Cell Biology

Article Title: Landscape expansion microscopy reveals interactions between membrane and phase-separated organelles

doi: 10.1083/jcb.202502035

Figure Lengend Snippet: land-ExM coupled with immunostaining LR-ExM for membrane-bound organelle visualization. (A–C) land-ExM total lipid (magenta) and protein (green) images of U2OS cells immunostained with anti-Tom20 antibodies (yellow). The anti-Tom20 antibodies are labeled LR-ExM second antibodies, which are second antibodies conjugated with NHS-digoxigenin-MA. Scale bar: 5 µm in pre-expansion unit. Linear expansion factor: 4. (D–G) Magnified images of A–C showing details of mitochondria. Scale bar: 1 µm in pre-expansion unit. (H) Intensity profile along the cyan line across the mitochondria in image (D). (I–K) land-ExM lipid (magenta) and protein (green) images of U2OS cells immunostained with anti-Sec61b antibodies (yellow). The anti-Sec61b antibodies are labeled LR-ExM second antibodies, which are second antibodies conjugated with NHS-digoxigenin-MA. Scale bar: 5 µm in pre-expansion unit. Linear expansion factor: 4. (L–O) Magnified in images of I–K showing details of ER. Scale bar: 1 µm in pre-expansion unit. (P) Intensity profile along the cyan line across the ER in image (L). All images were taken with an Airyscan microscope.

Article Snippet: Sec61b (D5Q1W) Rabbit mAb , Cell Signaling Technology , 14648.

Techniques: Immunostaining, Membrane, Labeling, Microscopy

Journal: The Journal of Cell Biology

Article Title: Landscape expansion microscopy reveals interactions between membrane and phase-separated organelles

doi: 10.1083/jcb.202502035

Figure Lengend Snippet: land-ExM reveals SGs at different locations of cells. (A–C) land-ExM images of U2OS cells untreated or treated with NaAsO2 for 20 or 60 min, then immunostained with anti-G3BP1 antibody. Scale bar: 5 µm in pre-expansion unit. Linear expansion factor: 4. (D–F) land-ExM images of U2OS cells stained with mCLING (magenta) and NHS ester dye (cyan) and immunostained with anti-G3BP1 (yellow) and anti-Sec61b (white) antibodies. Cells were untreated or treated with NaAsO2 for 20 min or 60 min. Scale bar: 5 µm in pre-expansion unit. Linear expansion factor: 4. (G) Magnified images of E showing SGs formed adjacent to ER (orange arrowheads). Scale bar: 1 µm in pre-expansion unit. (H) Analysis of the number of nuclear tunnels per cell with or without 60 min NaAsO2 treatment. Each bar represents the mean ± standard error of more than 18 cells. The ns indicates P > 0.05 by Welch’s t test. (I) Analysis of the diameter of nuclear tunnels in cells with or without 60 min NaAsO2 treatment. Each bar represents the mean ± standard error of more than 20 cells. ns indicates P > 0.05 by Welch’s t test. All images were taken with an Airyscan microscope. The cell shown in F is also shown in .

Article Snippet: Sec61b (D5Q1W) Rabbit mAb , Cell Signaling Technology , 14648.

Techniques: Staining, Microscopy

Journal: The Journal of Cell Biology

Article Title: Landscape expansion microscopy reveals interactions between membrane and phase-separated organelles

doi: 10.1083/jcb.202502035

Figure Lengend Snippet: The nuclear tunnel forms a triple-organellar contact site that includes the SG, the nucleolus, and itself. (A) land-ExM protein (gray) image of U2OS cells immunostained with anti-G3BP1 (red) antibody. Cells were treated with NaAsO 2 for 1 h. Scale bar: 5 µm in pre-expansion unit. Linear expansion factor: 4. (B–D) Different views of SG in the white dashed box of A. Scale bar: 1 µm in pre-expansion unit. (E) 3D rendering of SG in the white dashed box of A. In the reference grid, the spacing of major and minor tick marks is 0.5 and 0.1 µm in pre-expansion unit. (F) land-ExM protein (gray) and lipid (blue) image of U2OS cells immunostained with anti-G3BP1 (red) antibody. Cells were treated with NaAsO 2 for 1 h. Scale bar: 5 µm in pre-expansion unit. Linear expansion factor: 4. (G–I) Different views of SG in the white dashed box 1 of F. Scale bar: 1 µm in pre-expansion unit. (J) 3D rendering of SG in the white dashed box 1 of F. In the reference grid, the spacing of major and minor tick marks is 0.5 and 0.1 µm in the pre-expansion unit. (K) 3D rendering of SGs in the white dashed box 1–4 of F. In the reference grid, the spacing of major and minor tick marks is 0.5 and 0.1 µm in the pre-expansion unit. (L) land-ExM protein (gray) image of U2OS cells immunostained with anti-G3BP1 (red) and anti-Sec61b (yellow) antibodies. Cells were treated with NaAsO 2 for 1 h. Scale bar: 5 µm in pre-expansion unit. Linear expansion factor: 4. (M–O) Different views of SG in the white dashed box 1 of L. Scale bar: 1 µm in pre-expansion unit. (P) 3D rendering of SG in the white dashed box 1 of L. In the reference grid, the spacing of major and minor tick marks is 0.5 and 0.1 µm in pre-expansion unit. (Q) 3D rendering of SGs in the white dashed box 1–4 of L. In the reference grid, the spacing of major and minor tick marks is 0.5 and 0.1 µm in pre-expansion unit. (R) Pie chart of nuclear tunnels with or without SGs. Total tunnels analyzed: 114. (S) Pie chart of SG-filled nuclear tunnels that contact nucleoli versus those that do not. Total tunnel analyzed: 83. All images were taken with an Airyscan microscope. The cell shown in A, F, and L is also shown in .

Article Snippet: Sec61b (D5Q1W) Rabbit mAb , Cell Signaling Technology , 14648.

Techniques: Microscopy