HeLa cells stably expressing GalNAcT2-GFP were transfected with either control siRNA or siRab41(4) duplexes at a concentration of 200 nM. 96 h post initial transfection, cells were incubated with Cy3-SLTB at 4° C for 30 min and then transferred to 19.5° C, and incubated for 1 h to accumulate Cy3-SLTB in endosomes (A). Cells were then shifted to 37° C, permissive temperature for Cy3-SLTB transport from endosomes to Golgi apparatus, and incubated for 20 or 60 min before being fixed. At the end of 20-min (B) or 60-min chase (C), Cy3-SLTB accumulated in Golgi apparatus in both control and Rab41 knockdown cells, indicating that Rab41 knockdown has little effect on the transport of SLTB from endosome to Golgi apparatus. In the overlay images, distribution of Cy3-SLTB was shown in red, whereas the Golgi apparatus displayed by the expression of GalNAcT2-GFP was shown in green.