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. Author manuscript; available in PMC: 2013 Jul 4.
Published in final edited form as: Nat Cell Biol. 2013 Jan;15(1):17–27. doi: 10.1038/ncb2646

Figure 6. Dynamic MTs are required for apical relocation of aPKC and lumen formation.

Figure 6

(a) Time-course of polarity and lumen formation in 3D ICR acini embedded within BM-matrix and treated with paclitaxel (100 nM). Untreated acini (i, iii, v) developed apical lumens normally and MTs became orientated apico-basally. In contrast, MT stabilization (ii, iv, vi) impaired aPKC reorientation and lumen formation. Bar: 15μm.

(b) Nocodazole prevents lumen formation.

(ci) Schematic Z-view of polarized MECs in monolayer with apical tight junctions and basolateral integrins. (ii) Integrins are absent from top surface (extracellular domain antibody to β1; red) and apical membrane cannot internalize Tfr-488 from the media.

(d) MECs overlaid with BM matrix display β1-integrins at the top (i) schematic Z-view (ii) confocal view and TfR-488 internalization from the media. Bar: 8μm.

(e) Tfr488 uptake was quantified using a fluorescence plate reader. Histogram shows a single experiment representative of n=3. **p < 0.02; ***p< 0.05.

(f) Confluent monolayers of ICR MECs displaying apical tight junctions (i) were treated with DMSO (i, iii) or with paclitaxel (ii, iv) for 1h prior to BM-overlay (iii, iv). BM-overlay induced ZO1 tight junction disruption in DMSO treated cells but paclitaxel treatment prevented the disruption. Bar: 8μm.