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. 2011 Jan 7;88(1):99–105. doi: 10.1016/j.ajhg.2010.12.003

Figure 4.

Figure 4

Asn355Lys Atlastin-1 Exhibits Altered GTPase Activity and Disrupts ER Morphology when It Is Expressed in Cells

(A) Myc-tagged wild-type atlastin-1 or the indicated missense mutants were immunopurified from COS7 cells, and GTP hydrolysis was plotted as a function of time (top). Representative thin-layer chromatography plates show conversion of GTP to GDP (bottom). Error bars represent means ± standard deviation.

(B) COS7 cells were transfected with Myc-tagged wild-type or mutant atlastin-1 and immunostained for Myc-epitope (red) and β-tubulin (green). Merged images are at the right. DAPI nuclear staining is in blue. The scale bar represents 20 μm.

(C) Quantification of ER disruption (three trials of n = 100 cells per condition, means ± SD); ∗p < 0.01.

(D) COS7 cells transfected with Myc-tagged wild-type or Asn355Lys atlastin-1 were immunoblotted for Myc-epitope. Actin levels were monitored as a control for protein loading.