Volume 8 issue 9 September 2007
Nuclear transport of Kir/Gem requires specific signals, importin α5 and is regulated by calmodulin and predicted serine phosphorylations
RN Mahalakshmi, K Nagashima, MY Ng, N Inagaki, W Hunziker, and P Béguin

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Supplemental Figure 1: A. Effect of heat-shock on the subcellular localization of Kir/Gem W269G.
Hela cells expressing Myc-Kir/Gem W269G were exposed to 43°C (panel b) or kept at 37°C for 60 min (panel a) and processed for immunofluorescence microscopy using Myc antibodies. The fraction of transfected cells showing no, partial or complete nuclear localization of Kir/Gem was determined for each condition in 100 randomly chosen cells.

 B. Subcellular distribution of different combinations of Ser mutations in β-galactosidase fusion proteins containing WT or the indicated mutated forms of the C-terminal 42 amino acids of Kir/Gem (amino acids 253-295). COS-1 cells were transfected with cDNAs for the indicated fusion proteins then processed for immunofluorescence microscopy using an HA antibody to label the fusion protein as described in Fig. 3 and 5. The subcellular distribution (nuclear or diffused) was determined for each fusion protein in 100 randomly chosen cells. (*) As mentioned in the text (Fig. 3), the fusion protein including the unique W269G mutation displayed a nuclear staining for 50-60% of the transfected cells, whereas the remaining cell population showed a diffused pattern.

Figure 1 (.jpg)