Figure 5
From: Functional transformations of bile acid transporters induced by high-affinity macromolecules
Interaction of LHe-tetraD with IBABP and retro-translocation process of ASBT.
(a) The interaction between LHe-tetraD and IBABP along with ASBT in SK-BR-3 cells. To facilitate representation, the blots are cropped and the full-length blots are presented in the Supplementary Figure S7. (b) The colocalization of individually labeled LHe-tetraD (green) and IBABP (red) in SK-BR-3 cells (nucleus, blue). Scale bar indicates 10 μm. (c) PLA between IBABP and the biotin-labeled LHe-tetraD in the SK-BR-3-treated cells. Scale bar indicates 10 μm ASBT is stained as green. Arrowheads indicate the co-localization of LHe-tetraD/IBABP-PLA and ASBT in the membrane. Arrow indicates that the complexes of LHe-tetraD/IBABP (red dots) are being dissociated from ASBT in the cytoplasm. (d) SK-BR-3 cells were treated with LHe-tetraD for 30 min, washed and incubated for additional 15 min at 37°C in HBSS before staining. The extensive co-localization of individually labeled ASBT (green) and Rab11 (red), a marker of recycling endosome, indicated that free ASBTs were located in the recycling endosomes for membrane retro-translocation. Scale bar indicates 10 μm.
