Figure 3.
Disruption of actin and microtubule cytoskeleton enhances GTP availability and rates of NCT. (A) Representative ratiometric images (405 nm/488 nm) of BJ-5ta cells expressing either GEVALNull or GEVAL30 treated with cytochalasin B. (B) Quantification of GEVAL ratiometric signal (405 nm/488 nm) in BJ-5ta cells expressing GEVALNull or GEVAL30 treated with cytochalasin B (GEVALNull control and cytochalasin B, n = 121 and n = 135, respectively; GEVAL30 control and cytochalasin B, n = 117 and n = 130, respectively. Results are from two independent replicates. (C) Import and export rates of BJ-5ta cells expressing LINuS treated with cytochalasin B, latrunculin B, or nocodazole (control n = 65, cytochalasin B n = 35, latrunculin B n = 34, nocodazole n = 36). Results are from three independent replicates. (D) Quantification of GEVAL ratiometric signal (405 nm/488 nm) in MCF10A cells expressing GEVALNull or GEVAL30 treated with cytochalasin B (GEVALNull control and cytochalasin B, n = 101 and n = 123, respectively; GEVAL30 control and cytochalasin B, n = 134 and n = 157, respectively). Results are from three independent replicates. (E) Import and export rates of MCF10A cells expressing LINuS treated with cytochalasin B (control n = 60, cytochalasin B n = 60; results from three independent replicates). (F) Quantification of GEVAL ratiometric signal (405 nm/488 nm) in BJ-5ta cells expressing GEVALNull or GEVAL30 depleted of vimentin (GEVALNull siControl and siVimentin, n = 128 and n = 136, respectively; GEVAL30 siControl and siVimentin, n = 179 and n = 118, respectively). Results are from three independent replicates. (G) Import and export rates of BJ-5ta cells expressing LINuS depleted of vimentin (siControl n = 89, siVimentin n = 74; three independent replicates). Significance was calculated using unpaired t test (B and D–G), or one-way ANOVA with Dunnett’s post hoc (C). ns P > 0.05, P***<0.001, P****<0.0001. Scale bar 10 μm.

Disruption of actin and microtubule cytoskeleton enhances GTP availability and rates of NCT. (A) Representative ratiometric images (405 nm/488 nm) of BJ-5ta cells expressing either GEVALNull or GEVAL30 treated with cytochalasin B. (B) Quantification of GEVAL ratiometric signal (405 nm/488 nm) in BJ-5ta cells expressing GEVALNull or GEVAL30 treated with cytochalasin B (GEVALNull control and cytochalasin B, n = 121 and n = 135, respectively; GEVAL30 control and cytochalasin B, n = 117 and n = 130, respectively. Results are from two independent replicates. (C) Import and export rates of BJ-5ta cells expressing LINuS treated with cytochalasin B, latrunculin B, or nocodazole (control n = 65, cytochalasin B n = 35, latrunculin B n = 34, nocodazole n = 36). Results are from three independent replicates. (D) Quantification of GEVAL ratiometric signal (405 nm/488 nm) in MCF10A cells expressing GEVALNull or GEVAL30 treated with cytochalasin B (GEVALNull control and cytochalasin B, n = 101 and n = 123, respectively; GEVAL30 control and cytochalasin B, n = 134 and n = 157, respectively). Results are from three independent replicates. (E) Import and export rates of MCF10A cells expressing LINuS treated with cytochalasin B (control n = 60, cytochalasin B n = 60; results from three independent replicates). (F) Quantification of GEVAL ratiometric signal (405 nm/488 nm) in BJ-5ta cells expressing GEVALNull or GEVAL30 depleted of vimentin (GEVALNull siControl and siVimentin, n = 128 and n = 136, respectively; GEVAL30 siControl and siVimentin, n = 179 and n = 118, respectively). Results are from three independent replicates. (G) Import and export rates of BJ-5ta cells expressing LINuS depleted of vimentin (siControl n = 89, siVimentin n = 74; three independent replicates). Significance was calculated using unpaired t test (B and D–G), or one-way ANOVA with Dunnett’s post hoc (C). ns P > 0.05, P***<0.001, P****<0.0001. Scale bar 10 μm.

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