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. 2022 Jun 28;13:3701. doi: 10.1038/s41467-022-31282-8

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 7 — Scale bar indicates 2 µm and values represent mean ± S.D. a 1,6-hexanediol could dissolve the Lsm7 foci formed under 2-DG treatment but did not seemingly affect the Pab1 granules (dig, digitonin; dig+hex, digitonin plus 1,6-hexanediol). Values represent percentage of cells with Lsm7 foci or SGs. Four biologically independent experiments were examined and >200 cells were analyzed for each (unpaired two-tailed t-test). ****p < 0.0001, ns = 0.1063. b 1,6-hexanediol could block 2-DG-induced Lsm7 foci or Pab1 granule formation. (left, gray area). Once 1,6-hexanediol was washed out from the media, Lsm7 foci formed promptly while Pab1 granules formed afterward (right). Four biologically independent experiments were examined and >200 cells were analyzed for each. c Dissolved foci phenotype of Pab1-RFP and Lsm7-GFP pre-treated with 1,6-hexanediol and digitonin (30 min) followed by addition of 2-DG for 2 h. The SG and Lsm7 foci formation was hampered in the WT Lsm7-GFP (left) strain as well as the lsm7∆90-103-GFP mutant (right). Three biologically independent experiments were examined and >200 cells were analyzed per clone. Values represent percentage of cells with Lsm7 foci or SGs. d Images depicting in vivo FRAP analysis on Lsm7-GFP, Pab1-RFP, and Dcp2-GFP foci after 2 h 2-DG treatment. Images display foci before, immediately after and 10 s after photobleaching. Representative images from three (Lsm7), six (Pab1), and five (Dcp2) independent experiments. e FRAP recovery curves for Lsm7-GFP, Pab1-RFP, and Dcp2-GFP foci after 2 h 2-DG stress. Since the recovery of Lsm7 foci was quick, the x-axis (time, seconds) is shorter than for Pab1-RFP and Dcp2-GFP. Three (Lsm7), six (Pab1), and five (Dcp2) independent experiments. f FRAP results depicting time (s) to full recovery after bleaching (left) and calculated half-time rate (right). Three (Lsm7), six (Pab1), and five (Dcp2) independent experiments (one-way ANOVA followed by Dunnett’s T3 multiple comparisons test, with individual variances computed for each comparison). Left to right: **p = 0.0035, **p = 0.0029, *p = 0.0134, ns = 0.0912. Source data are provided as a Source Data file.