Fig. 7

CI-M6PR sorting to endosome carriers is impaired without Rab4b. Internalization of CI-M6PR (iCI-M6PR) is tracked by fluorescently coupled anti-CI-M6PR antibodies using protocol 2 (Additional file 1: Fig. S2A, 10 min chase instead of 30 min) in cells treated with control or anti-Rab4b siRNA. A Representative images after the 10 min chase (iCI-M6PR in 3D, after its volume reconstruction, VPS52 converted to spot, and VPS52 spot close to iCI-M6PR volume)The bar is for 20 µm. Image processing is described in the Material and Methods section. The same threshold and the same filters were applied for all the images. B Quantification of the number of VPS52 spots near the iCI-M6PR volume in cells treated with control siRNA (filled circles) and anti-Rab4b siRNA (empty circles) before and after the 10 min chase. **, *** indicate that the differences are significant with p < 0.01 and p < 0.001, respectively (one-way ANOVA)