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Fig. S1
. Zebrafish Stmn4 was localized to Golgi apparatus in a cysteine-dependent manner.
(a) Construct design: the mCherry with or without a C terminal full-length coding sequence of stmn4 gene, containing two N terminal domains (A and A') and a stathmin-like domain was cloned into the pCS2+ vector. Two cysteine sites, C20 and C22, on the A domain were changed to alanine that could interfere with Golgi membrane-tethering of Stmn4. (b) Hela cells were co-transfected with Golgi-YFP to reveal Golgi in green and mCherry containing pCS2+ vector without or with wildtype or Cysteine-mutated stathmin as indicated in red. Cells were imaged and photographed under confocal microscopy and the overlapping signal in yellow indicated the stathmin was located in Golgi apparatus. A higher magnification of a dashed area is shown at the lower right corner for each panel. Scale bars are shown only on the upper left panel. To analyze the localization of Stmn4 in Golgi apparatus, the overlapping yellow signals were changed to white for clarity using the ImageJ software as shown in the higher magnification graphs. The correlations between treatments for the Golgi localization of stathmin are shown by calculating their Pearson’s correlation coefficients and statistically analyzed by one-way ANOVA as resented in the lower right graph. Data represents mean + s.e.m. *P<0.05.