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. 2018 Aug 21;7:53–67. doi: 10.1016/j.isci.2018.08.014

Figure 1.

Figure 1

Caveolin-1 Is Strongly Expressed in the Immature Cortical Neurons and Required for Clathrin-Independent Endocytosis in Neurons

(A) Immunoblot analyses of lysates from the indicated tissues at E15 with the indicated antibodies. The ratios of immunoblot band intensities of caveolin-1/β-actin, as determined by Las-3000mini (Fuji-film), are indicated. See also Figure S1.

(B–D) Cryosections of cerebral cortices at E17 were immunostained with anti-caveolin-1 (B and C) or anti-α-adaptin (D) and DAPI (nuclear staining). (C) High magnification images near the lower intermediate zone. White arrows indicate immature neurites. MZ, marginal zone; CP, cortical plate; IZ, intermediate zone; VZ, ventricular zone. See also Figure S2.

(E) Primary cortical neurons from E15 cerebral cortices incubated for 2 days in vitro and stained with anti-GD3 (green) and anti-caveolin-1 (red) antibodies. Anti-GD3 antibody may induce the clustering of GD3 after fixation (because lipids are not fixed by 4% PFA), and a part of caveolin-1 is also observed in the GD3-containing small aggregates on the membrane, suggesting that caveolin-1 may interact with GD3 and form the co-aggregates with GD3 after fixation. See also Figure S3.

(F) Primary cortical neurons from E15 cerebral cortices incubated for 2 days in vitro and stained with anti-CD71 (green) and anti-caveolin-1 (red) antibodies. See also Figure S3.

(G) Immunoblot analyses of lysates from the indicated tissues (E15 or adult mice) with anti-Cavin-1 (black arrow) and anti-β-actin antibodies.

(H and I) (H) Primary cortical neurons from E15 cerebral cortices were transfected with CAG = ECFP-Mem (blue, a marker for transfected cells), incubated for 2 days in vitro, and treated with BODIPY-LacCer for 30 min before fixation. White arrows indicate the perinuclear accumulation of BODIPY-LacCer (green). The graph in (I) shows the ratio of cells with perinuclear accumulation of BODIPY-LacCer. n = 3 (control: 82 cells, Cav1-sh490: 87 cells).

Each score represents the mean of ratios ± SEM. Significance compared with control was determined by Student's t test (p = 0.0142) and Mann-Whitney's U test (p = 0.0495). *p < 0.05. See also Figures S4 and S5. Scale bars: 100 μm in (B), 10 μm in (C), 100 μm in (D), 2 μm in (E and F), and 3 μm in (H).