Figure 2.

CCL21-Positive Vesicles Localize to Golgi-Secretory Pathway and Traffic along Microtubules

(A) Giantin (white) and nuclear staining (DAPI, blue) of CCL21-mCherry (red), CCL21ΔN-mCherry, or CCL21ΔC-mCherry expressing LECs. Insets show Golgi localized mCherry signal (yellow arrows) with or without staining of Golgi marker giantin.

(B) Quantification of mCherry intensities in supernatants of CCL21-mCherry, CCL21ΔN-mCherry, and CCL21ΔC-mCherry expressing LECs. Dot blot graph shows a mean ± SD of pooled data from two independent experiments, n = 4 for each construct. Intensities are normalized to media background, which is set as 1.

(C) Basolateral non-directed tracks of TIRF-imaged CCL21-mCherry-positive vesicles are indicated with white, directed tracks heading toward Golgi area with green, directed tracks toward cell periphery with red, the Golgi apparatus with a dashed yellow line, and the boundaries of cell contact surface with dashed white line. Yellow arrow indicates a white trajectory representing random movement within a confined area.

(D) TIRF imaging of CCL21ΔC-mCherry (red) and EGFP-tubulin-α (green) expressing primary LEC. White arrow highlights one CCL21ΔC-mCherry positive vesicle (red) throughout the image series. See Movie S1. Scale bars, 10 μm (A); 2 μm (C and D).

See also Figure S2.