Anti-Ultraspiracle抗体
Anti-Ultraspiracle antibody
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(2 Publications)
Rabbit Polyclonal USP antibody. Suitable for ICC, WB and reacts with Drosophila melanogaster samples. Cited in 2 publications. Immunogen corresponding to Synthetic Peptide within Silkworm USP.
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Cf1, NR2B4, CG4380, usp, Protein ultraspiracle, Chorion factor 1, Nuclear receptor subfamily 2 group B member 4, XR2C
- WB
CiteAb
Western blot - Anti-Ultraspiracle antibody (AB106341)
Western Blotting using Anti-Ultraspiracle antibody, ab106341. Publication image from Brehm, A. et al., 2017, Nat Commun, 28378812. Legend direct from paper.
dMi-2 forms a complex with EcR and competes with USP for binding to EcR.(a) dMi-2 and EcR interact. Nuclear extracts from untreated and ecdysone exposed (+20HE) S2 cells were immunoprecipitated with dMi-2 antibody, dp53 antibody or IgG as indicated on top (lanes 2–4 and 6–8). 1% input was loaded in lanes 1 and 5. Antibodies used for western blot analysis are indicated on the right, molecular masses on the left. (b) Sf9 cells were infected with recombinant baculoviruses directing the expression of dMi-2-FLAG, dMi-2 or EcR-FLAG as indicated on top. Extracts were immunoprecipitated with FLAG antibody and washed with high salt buffer. Immunoprecipitates were then analysed by SDS–PAGE and Coomassie staining. Lane 1 : molecular weight marker. Lanes 2, 4 and 6 : 500 ng protein, lanes 3, 5 and 7 : 1 µg protein. (c) dMi-2 and USP bind EcR in a mutually exclusive manner. Sf9 cells that were either left untreated (left panels) or were exposed to ecdysone (+20HE, right panels) were infected with recombinant baculoviruses expressing dMi-2-FLAG, EcR or HA–USP as indicated on top. Extracts were analysed by western blot for expression of recombinant proteins (top panels). Extracts were immunoprecipitated with FLAG (FLAG IP, middle panels) or HA (HA IP, bottom panels) antibody and immunoprecipitates were analysed by western blot. Recombinant proteins detected by western blots are indicated on the right.
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- WB
CiteAb
Western blot - Anti-Ultraspiracle antibody (AB106341)
Western Blotting using Anti-Ultraspiracle antibody, ab106341. Publication image from Brehm, A. et al., 2017, Nat Commun, 28378812. Legend direct from paper.
EcR but not USP is essential for recruitment of dMi-2 to ecdysone-activated genes.(a) Enrichment of predicted EcR–USP-binding sites in EIMRs. The histogram shows the distribution (sampled from 10,000 runs) of putative EcR–USP site number within the same number of randomly selected genomic regions with the same sizes as the experimentally determined EIMRs. The red line indicates the number of computationally identified EcR–USP-binding motifs within EIMRs (142; P<10-8). The P-value for the observed number of binding motifs was estimated from the normal distribution that can be approximated based on the displayed simulated distribution, that is, using mean and s.d. estimates from the simulated sample. (b) Western blot analysis of protein extracts of S2 cells treated with dsRNA directed against GFP (control), EcR or USP. Cells were either left untreated (−20HE) or were treated with ecdysone (+20HE). Antibodies used are shown on the right, molecular masses on the left. Tubulin served as a loading control. (c) dMi-2 binding to Br-C and vrille following EcR and USP depletion analysed by ChIP-qPCR. Chromatin was prepared from S2 cells that were first treated with dsRNA against GFP (control), EcR or USP and then either left untreated or were exposed to 1 µM ecdysone for 6 h as indicated. ChIP-qPCR was performed with dMi-2 antibody. Error bars denote s.d. of technical triplicates. Experiments were performed as biological triplicates. One representative example is shown.
false
- WB
CiteAb
Western blot - Anti-Ultraspiracle antibody (AB106341)
Western Blotting using Anti-Ultraspiracle antibody, ab106341. Publication image from Brehm, A. et al., 2017, Nat Commun, 28378812. Legend direct from paper.
dMi-2 constrains the transcription of ecdysone-activated genes and contributes to a closed chromatin structure.(a) S2 cells were RNAi-depleted of GFP (control), dMi-2, EcR or USP and either left untreated (−20HE) or were treated with 1 µM ecdysone for 6 h (+20HE). Nuclear extracts were prepared and analysed by western blot. Asterisk (*) denotes USP antibody-crossreactive protein. Tubulin served as loading control. Representative examples of three independent RNAi experiments are shown. (b) RT–qPCR analysis of dMi-2 (top), EcR (middle) and USP (bottom) expression. RNA levels in control cells (GFP RNAi) were set to 1 and RNA levels in other RNAi-treated cells are depicted relative to the level in control cells. (c) Time course of Br-C and (d) vrille RNA expression over 6 h of ecdysone treatment. Before ecdysone addition, cells were RNAi-depleted of GFP (control), dMi-2, EcR or USP as indicated. RNA levels were determined by RT–qPCR relative to rp49 and adjusted to 100 at the latest time point of control cells (GFP RNAi). Other ratios were expressed relative to this. Data shown are mean value±s.e.m. of three independent experiments. (e) Derepression of ecdysone-regulated genes after dMi-2, EcR and USP depletion. S2 cells were RNAi-depleted of GFP (control), dMi-2, EcR or USP. Cells were then left untreated or were treated for 6 h with 1 µM 20HE as shown. Data shown are mean value±s.e.m. of two independent experiments. (f) Schematic representation of amplimer positions (A–G) within the vrille locus. Note that regions A and D contain EIMRs (see Fig. 1e). (g) Cells were RNAi-depleted of GFP (control) or dMi-2 and then left untreated or treated with ecdysone (+20HE). Chromatin was prepared, digested with MNase and amplified by qPCR. Relative MNase protection was calculated and plotted (see Methods section). The sample with the highest MNase protection value for each genomic region was set to 1. Error bars denote s.d. of technical triplicates. Experiment was performed as biological triplicates, one representative experiment is shown.
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补充信息
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
The ecdysone receptor complex including Ultraspiracle has a significant role in the process of insect molting and metamorphosis. Ultraspiracle as part of this heterodimer with EcR binds to ecdysone response elements on DNA leading to the activation of specific genes necessary for development. This interaction influences various biological processes such as cell cycle regulation and programmed cell death which are important during the transition from larva to pupa and finally to an adult insect.
Pathways
The ecdysone receptor complex involves Ultraspiracle and EcR in the ecdysone signaling pathway which regulates insect development and metamorphosis. The pathway is key in controlling the expression of genes that determine molting and differentiation during developmental stages. USP and EcR interact with other transcription factors and co-regulators to fine-tune the gene expression profiles necessary for these processes. Additionally interactions with proteins like Heat Shock Protein 90 (Hsp90) assist in stabilizing the ecdysone receptor complex ensuring proper signaling.
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文献 (2)
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PLoS genetics 13:e1006788 PubMed28493870
2017
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Unspecified application
Species
Unspecified reactive species
Nature communications 8:14806 PubMed28378812
2017
Applications
WB
Species
Unspecified reactive species
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