Export of cyst wall material and Golgi organelle neogenesis in Giardia lamblia depend on endoplasmic reticulum exit sites
Version of Record online: 2 DEC 2012
© 2012 Blackwell Publishing Ltd
Volume 15, Issue 4, pages 537–553, April 2013
How to Cite
Faso, C., Konrad, C., Schraner, E. M. and Hehl, A. B. (2013), Export of cyst wall material and Golgi organelle neogenesis in Giardia lamblia depend on endoplasmic reticulum exit sites. Cellular Microbiology, 15: 537–553. doi: 10.1111/cmi.12054
- Issue online: 14 MAR 2013
- Version of Record online: 2 DEC 2012
- Accepted manuscript online: 25 OCT 2012 02:05AM EST
- Manuscript Accepted: 11 OCT 2012
- Manuscript Revised: 5 OCT 2012
- Manuscript Received: 11 MAY 2012
- Swiss National Science Foundation. Grant Number: 31003A_125389
Fig. S1. Myc-Sar1 GTPase detection in cell lines mycSar-Sec23HA and mycSarmut-Sec23HA. Representative images for Sec23HA and Sar1 GTPase deposition in cell lines mycSar-Sec23HA and mycSarmut-Sec23HA, using wide-field fluorescence microscopy. Sec23HA accumulation at ERES is not affected by overexpression of wild-type Sar1 GTPase (A) while it is clearly disrupted in the presence of mutated Sar1 GTPase (C). Both wild-type (B) and mutant (D) myc-tagged Sar1 GTPase show a diffused localization pattern. Nuclei were labelled with DAPI (blue). Insets: bright-field images. Scale bar: 1 μm. A SAG1 construct carrying a VSP trafficking signal and Sec23HA colocalize at ERES. (E) Representative image acquired by wide-field fluorescence microscopy of differentiating trophozoites expressing construct SAG1-VSPct. The SAG1 exodomain (red) was detected at the plasma membrane and at discrete intracellular sites reminiscent of ERES. Nuclei were labelled with DAPI (blue). Inset: bright-field image. Scale bar: 1 μm. (F) Representative image of a differentiating trophozoite coexpressing SAG1-VSPct (red) and Sec23HA (green) at ERES, acquired by wide-field fluorescence microscopy. Nuclei were labelled with DAPI (blue). Inset: bright-field image. Scale bar: 1 μm. (G) Confirmation of Gl-TPI-directed chromosomal integration of pPACV-Integ-derived constructs in cell lines Sec23HA, mycSec24-Sec23HA, mycSar-Sec23HA and mycSarmut-Sec23HA. Genomic DNA samples from untransfected cells (lane 1) and from cell lines Sec23HA (lane 2), mycSec24-Sec23HA (lane 3), mycSar-Sec23HA (lane 4) and mycSarmut-Sec23HA (lane 5) all present the control amplicons (c. 1100 bp) derived from a non-targeted locus. However, only samples derived from stably transfected lines (lanes 2–5) present the integration-derived c. 2000 bp amplicon, indicating that construct integration in gDNA of stably transfected lines occurred within the Gl-TPI locus. M, GeneRuler DNA ladder marker (Fermentas); selected band sizes in bp are indicated on the left.
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