Figure 4. CAV1 and cavin-1 in the... | Rockefeller University Press

Figure 4.

$Figure 4. CAV1 and cavin-1 in the endosomal pathway. (A–C) CAV1-mCherry localized to Rab5-positive EE and to Rab7- and Lamp1-positive LE/LYS. Single confocal sections of fixed (A) or living (B and C) cells were acquired 12 h after transfection. (bottom) Insets show enlargements of boxed areas. (D) Enlarged views of individual organelles. In EE, CAV1-mCherry was present in the limiting membranes and, in LE/LYS, in the lumen of the organelles. (E and F) Cavin-1–mCherry localized to Rab5-positive EE but not to Rab7-positive LE. Cavin-1–mCherry was coexpressed with GFP-tagged endosomal makers and CAV1-HA (not depicted), and images acquired 12 h after transfection form living cells using an epifluorescence setup. (bottom) Insets show enlargements of the boxed areas. Arrowheads point to endosomal organelles indicating presence (A–C and E) or absence (F) of colocalization between markers. (G) Colocalization analysis of CAV1-mCherry endosomal structures with endosomal markers. The fraction of CAV1-mCherry endosomes per cell colocalizing with EGFP-Rab5 (n = 68 cells), EGFP-Rab7 (n = 68 cells), and Lamp1-EGFP (n = 43 cells) was determined as detailed in Materials and methods (Fig. S2 A) from images acquired from living cells using an epifluorescence setup. Box plots show medians, lower and upper quartiles (line and box), 10th and 90th percentiles (whiskers), and outliers (•). Bars: (A–C, E, and F) 10 µm; (D) 1 µm.$

CAV1 and cavin-1 in the endosomal pathway. (A–C) CAV1-mCherry localized to Rab5-positive EE and to Rab7- and Lamp1-positive LE/LYS. Single confocal sections of fixed (A) or living (B and C) cells were acquired 12 h after transfection. (bottom) Insets show enlargements of boxed areas. (D) Enlarged views of individual organelles. In EE, CAV1-mCherry was present in the limiting membranes and, in LE/LYS, in the lumen of the organelles. (E and F) Cavin-1–mCherry localized to Rab5-positive EE but not to Rab7-positive LE. Cavin-1–mCherry was coexpressed with GFP-tagged endosomal makers and CAV1-HA (not depicted), and images acquired 12 h after transfection form living cells using an epifluorescence setup. (bottom) Insets show enlargements of the boxed areas. Arrowheads point to endosomal organelles indicating presence (A–C and E) or absence (F) of colocalization between markers. (G) Colocalization analysis of CAV1-mCherry endosomal structures with endosomal markers. The fraction of CAV1-mCherry endosomes per cell colocalizing with EGFP-Rab5 (n = 68 cells), EGFP-Rab7 (n = 68 cells), and Lamp1-EGFP (n = 43 cells) was determined as detailed in Materials and methods (Fig. S2 A) from images acquired from living cells using an epifluorescence setup. Box plots show medians, lower and upper quartiles (line and box), 10th and 90th percentiles (whiskers), and outliers (•). Bars: (A–C, E, and F) 10 µm; (D) 1 µm.