Figure 7.
POM121 depletion relieves NE abnormalities. (A) Confocal images of 4TorKO cells transfected with MLF2-GFP and treated with 100 nM nontargeting (NT) siRNA, ELYS siRNA, or POM121 siRNA. Cells were counterstained with anti–K48-Ub (red) antibody. Scale bar, 10 µm; inset, 2 µm. (B) The relative amounts of ELYS and POM121 mRNA transcripts following 50 nM or 100 nM siRNA treatments were analyzed by RT-qPCR with the indicated primers. Relative mRNA levels were normalized to the siNT control. The values are means of three independent replicates (n = 3), and error bars indicate ±SD. (C) Graph of the percentage of cells with MLF2-GFP and K48-Ub foci after treatment with 100 nM of the indicated siRNAs. Each result is the average of three independent experiments with 75 cells each. The standard error of the mean is indicated by the error bars. P value was calculated using a two-tailed t test. (D) Graph of the number of blebs observed per EM cross section upon treatment with 100 nM of the indicated siRNA. Each point shows the number of blebs per 30 µm of NE in an individual cross section. The average blebs per 30 µm of NE is shown by the bar graph, and the standard error of the mean is shown in the error bars. 30 cells from each sample were counted. P value was calculated using a two-tailed t test. (E) EM images of 4TorKO control cells or cells treated with 100 nM siRNA targeting ELYS or POM121. The bottom panel is an enlarged view of the boxed region in the top panel. Scale bars, 500 nm (top panels) and 250 nm (bottom panels). N, nucleus; C, cytoplasm.

POM121 depletion relieves NE abnormalities. (A) Confocal images of 4TorKO cells transfected with MLF2-GFP and treated with 100 nM nontargeting (NT) siRNA, ELYS siRNA, or POM121 siRNA. Cells were counterstained with anti–K48-Ub (red) antibody. Scale bar, 10 µm; inset, 2 µm. (B) The relative amounts of ELYS and POM121 mRNA transcripts following 50 nM or 100 nM siRNA treatments were analyzed by RT-qPCR with the indicated primers. Relative mRNA levels were normalized to the siNT control. The values are means of three independent replicates (n = 3), and error bars indicate ±SD. (C) Graph of the percentage of cells with MLF2-GFP and K48-Ub foci after treatment with 100 nM of the indicated siRNAs. Each result is the average of three independent experiments with 75 cells each. The standard error of the mean is indicated by the error bars. P value was calculated using a two-tailed t test. (D) Graph of the number of blebs observed per EM cross section upon treatment with 100 nM of the indicated siRNA. Each point shows the number of blebs per 30 µm of NE in an individual cross section. The average blebs per 30 µm of NE is shown by the bar graph, and the standard error of the mean is shown in the error bars. 30 cells from each sample were counted. P value was calculated using a two-tailed t test. (E) EM images of 4TorKO control cells or cells treated with 100 nM siRNA targeting ELYS or POM121. The bottom panel is an enlarged view of the boxed region in the top panel. Scale bars, 500 nm (top panels) and 250 nm (bottom panels). N, nucleus; C, cytoplasm.

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