Zhang, et al (2003): Figure 4

Supplementary Material From Singer Lab Publications:

Active transport of the survival motor neuron protein and the role of exon-7 in cytoplasmic localization

J Neurosci 23(16):6627-6637 (2003 July 23)

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Figure 4. Association of EGFP-SMN with the actin cytoskeleton in fibroblasts.

A, EGFP-SMN was distributed predominantly in the cytoplasm and signal often accumulated at the leading edge of cultured fibroblast (arrow). One or two dots were also observed in the nucleus.

B, Corresponding DIC image.

C, EGFP-SMN granules were retained after Triton X-100 extraction in buffer (arrow).

D, DIC image.

E, Retention of EGFP-SMN on the cytoskeleton after colchicine treatment and Triton X-100 extraction (arrows).

F, This procedure did not disrupt actin filaments, although microtubules were depolymerized (data not shown).

G, In contrast, cytochalasin-D and Triton X-100 extraction released EGFP-SMN, and little signal was apparent.

H, Note the disruption of most actin filaments after cytochalasin-D treatment.

I, Quantitative analysis of the mean cytoplasmic fluorescence intensity in Triton X-100 extracted cells demonstrated a 70% reduction after cytochalasin-D treatment and a 26% reduction after colchicine treatment.