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. 2020 Jul 7;48(15):8332–8348. doi: 10.1093/nar/gkaa552

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© The Author(s) 2020. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

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Figure 3. — Conserved regions of NC2 subunits are required for transcriptional activation of cat-3. (A, B) Schematic drawing of N. crassa NC2α (A) and NC2β (B) subunits and their various deletion mutants. The position of the histone fold (red rectangle), acidic (green rectangle), TBP-binding (blue rectangle) and repression (gray rectangle) regions are indicated. (C) Plate assays analyzing mycelial growth of different deletion strains across NC2α coding region at endogenous locus on plates with 0 or 10 mM H₂O₂. Cultures were inoculated in plates at 25°C under constant light. (D) In-gel assay analysis of the CAT-3 activity levels in different deletion strains across NC2α coding region at endogenous locus. (E) Western blot analyses showing the levels of CAT-3 protein in different deletion strains across NC2α coding region at endogenous locus. The membrane stained by coomassie blue represents the total protein in each sample and acts as loading control for western blot. (F) RT-qPCR assays analyzing the levels of cat-3 mRNA in different deletion strains across NC2α coding region at endogenous locus. Error bars show s.d. (n = 3). Significance was evaluated by using a two-tailed t-test. **P < 0.01 and ***P < 0.001 versus WT. (G) Plate assays analyzing mycelial growth of different deletion strains across NC2β coding region at endogenous locus on plates with 0 or 10 mM H₂O₂. Cultures were inoculated in plates at 25°C under constant light. (H) In-gel assay analysis of the CAT-3 activity levels in different deletion strains across NC2β coding region at endogenous locus. (I) Western blot analyses showing the levels of CAT-3 protein in different deletion strains across NC2β coding region at endogenous locus. The membrane stained by coomassie blue represents the total protein in each sample and acts as loading control for western blot. (J) RT-qPCR assays analyzing the levels of cat-3 mRNA in different deletion strains across NC2β coding region at endogenous locus. Error bars show s.d. (n = 3). Significance was evaluated by using a two-tailed t-test. **P < 0.01 and ***P < 0.001 versus WT.