Phylogenetic tree of the TRAFs and diagrams of their protein domain structure

<p><b>Copyright information:</b></p><p>Taken from "Conservation and divergence of gene families encoding components of innate immune response systems in zebrafish"</p><p>http://genomebiology.com/2007/8/11/R251</p><p>Genome Biology 2007;8(11):R251-R251.</p><p>Published online 27 Nov 2007</p><p>PMCID:PMC2258186.</p><p></p> Details are as in Figure 2, except that the scale shows 0.2 amino acid substitutions. TRAF, tumor necrosis factor receptor-associated factor

Slik activates Moesin at the luminal membrane in terminal branches.

<p>(A–A″) Region of a terminal branch from a third intar larva showing Slik (A) and activated pMoesin (A′) staining. Slik colocalizes with pMoesin at the luminal membrane (A″). The terminal cell is labeled with cytoplasmic GFP, pseudo-colored in blue (A″). (B–C″) pMoesin staining in control (B′) and Slik-depleted (C′) terminal cells. No pMoesin is seen at the luminal membrane in Slik-depleted cells (C′). (D–E″) Higher magnification of a control cell (E) and an example of a <i>slik</i>-RNAi expressing terminal cell (D) with residual Slik staining but no pMoesin. (F–G″) In <i>slik</i>¹ homozygous mutant embryos pMoesin staining is present in some branches (F′, green arrowhead) and reduced or absent (F′, white arrowhead) in others. The tracheal tubes are labeled with CBD-Alexa 633 (F′, F″, G and G″, pseudo-colored, in blue). The larval terminal cell is labeled with cytoplasmic GFP, pseudo-colored in blue (D″ and E″). A–C″ and F–G″ are projections of confocal image stacks. D–E″ are single focal planes from image stacks. Scale bars: (A–A″ and D–E″) 5 µm, (B–C″) 20 µm, (F–G″) 20 µm.</p

Slik is necessary for terminal cell development.

<p>(A–D) The terminal cells are labeled with cytoplasmic GFP (green). Examples of control (A), <i>slik</i>¹ mutant (B, C) and <i>slik</i>-RNAi (D) expressing terminal cells in third instar larvae. (E) <i>slik</i>¹ mutant and <i>slik</i>-RNAi expressing terminal cells have a reduced number of branches (P value <0.0001 by two tailed T test) compared to control cells. (G–J) Slik is expressed and is apically enriched in tracheal cells during different stages of embryonic development. Tracheal cells (H, I and J) are labeled by immunostaining for Dof. (J′) Slik expression in third instar larval terminal branch labeled with cytoplasmic GFP. Slik is distributed in the cytoplasm with an enrichment at the luminal membrane (J′). A–D and G–I″ are projections of confocal image stacks. (J–J″) is a single focal plane from a confocal image stack. All constructs (GFP, RNAi) are expressed under the control of the <i>btl</i>-GAL4, UAS-GFP transgene (control). Scale bars: (A–B) 50 µm, (G–I″) 10 µm, (J–J″) 5 µm.</p

Breathless regulates pMoesin at the luminal membrane in terminal branches.

<p>(A–D″) pMoesin staining in control terminal cells (A–A″), and terminal cells expressing <i>btl</i>-RNAi (B-B″), <i>slik</i>-RNAi (C–C″) or <i>egfr</i>-RNAi (D–D″). pMoesin staining is absent in <i>btl</i>-RNAi (B′) and in <i>slik</i>-RNAi cells (C′). In <i>egfr</i>-RNAi cells (D′) pMoesin is present at levels comparable to control cells (A′). In both Btl and EGFR-depleted terminal cells branch numbers are significantly reduced (E, P value <0.0001 by two-tailed T test in both cases). In 63% of Btl-depleted cells (N = 19) pMoesin was absent, 27% showed pMoesin staining. Genotypes of crosses and number of terminal cells scored: Blue (control): <i>btl</i>-GAL4, UAS-GFP (N = 11). Red: <i>btl</i>-RNAi (N = 11). Violet: <i>egfr</i>-RNAi (n = 10). A–D″ are projections of confocal image stacks. Scale bars: (A–D″) 25 µm.</p

Depletion Moesin does not suppress the FGF signaling overactivation phenotype in terminal cells.

<p>(A–B″) Total Moesin staining in larvae expressing constitutively active FGFR alone (λ<i>btl</i>, A′) or in combination with <i>moe</i>-RNAi (B′) Moesin staining is detectable in the control (A′) but not in <i>moe</i>-RNAi; λ<i>btl</i> cells(B′). (C–D″) pMoesin staining in larvae expressing constitutively active FGFR alone (λ<i>btl</i>, C) or in combination with <i>moe</i>-RNAi (D′). pMoesin is absent or reduced in the <i>slik</i>-RNAi; λ<i>btl</i> cells (D′). A–D″ are projections of confocal image stacks. Scale bars: (A–D″) 40 µm.</p

Overactivation of FGF signaling overrides requirement for Slik in terminal cell branching.

<p>(A–D) Moesin (A, B) and Slik (C–D) staining in larvae expressing constitutively active FGFR alone (λ<i>btl</i>, A′) or in combination with <i>slik</i>-RNAi (B′). λ<i>btl</i>, induces excessive branching; pMoesin and Slik staining are detectable in the λ<i>btl</i> expressing cells, (A′, C′) but absent if <i>slik</i>-RNAi is co-expressed(B′, D′). Depletion of Slik in λ<i>btl</i> expressed terminal cells does not affect expression or membrane localization of total Moesin (E and F′). A–E″ are projections of confocal image stacks. F–F″ are single focal planes from a image stack. Scale bars: (A–E″) 30 µm, (F–F″) 5 µm.</p

Breathless-mediated regulation of Moesin is specific for terminal cells. (A–B″) total Moesin in control (A′) and <i>btl</i>-RNAi (B′) expressing terminal cells.

<p>(C–D″) Slik staining in control (C′) and <i>btl</i>-RNAi (D′) expressing terminal cells. Depleting Btl does not affect total Moesin (B′) or Slik (D′) in terminal cells. (E–F″) pMoesin staining in control and <i>btl</i>^LG19 mutant embryos. pMoesin localizes apically in control (E′) and <i>btl</i>^LG19 (F′) mutant embryonic tracheal branches. The insets in F′ and F″ are zoomed images from the boxed regions. (G–I″) pMoesin staining in control (G), <i>btl-</i>depleted (H) and <i>slik</i>-depleted (I) larval dorsal trunk, tracheoblasts and fusion cells (boxed regions in G′ and H′). pMoesin staining in <i>btl-</i>depleted cells is comparable to the control, whereas it is absent in <i>slik</i>-depleted cells. A-I″ are projections of confocal image stacks. Scale bars: (A–D″) 50 µm, (E–I″) 25 µm.</p

Estimating the growth of preprint review over time.

Preprints evaluated per month on Sciety, excluding reviews conducted by automated tools (ScreenIT) and reviews by journals posted after publication of the journal version (source data available [20]). This chart includes data from the following services, regardless of which server the preprints they evaluate have been posted to: eLife, Review Commons, Arcadia Science, preLights, Rapid Reviews, PREreview, NCRC, Peer Community In (Evolutionary Biology, Ecology, Zoology, Animal Science, Neuroscience, Paleontology, Archaeology), PeerRef, Biophysics Colab, ASAPbio (and ASAPbio-SciELO) crowd review, Life Science Editors (including Foundation), and The Unjournal. Data have been collected and provided by Sciety. Reviews posted to comment sections of preprint servers are not included, and depending on the policies of individual services, some of the evaluations included in this chart may not meet our definition of preprint review.</p