Nup133 is required for nuclear pore basket assembly and dynamics in embryonic stem cells. B. Souquet et al.
Contributors: Valérie DOYE, Alessandro Berto
... Pull-down experiments using as bait Nup153-NTD[aa1-245](negative control) or [aa1-339] and whole cell extracts from WT (HM1) or Nup133-/- (KO, #14) mESCs. Inputs (1x equivalent) and eluates (64x equivalent, dataset (i) or 48x equivalent, dataset (ii)) were analyzed by western blot. These western blots revealed a similar enrichment of Tpr and Y-complex proteins (Nup107,Nup96, Nup85) in Nup153-NTD [aa 1-339]-purified extracts from either WT or Nup133-/- mESCs. Anti-Nup98 and GAPDH were used as negative controls. The zip-file includes 2 folders correspond to datasets (i): Souquet et al Figure 4D (i, 64x eluates) and (ii): Souquet et al Figure 4D (ii, 48x eluates), respectively. In the dataset (ii), 3 independently processed eluates were analyzed. Each folder encompasses a general montage, in which the areas used for Figure 4C are indicated in red, as well as 5 sub-folders corresponding to the original 16-bit images (each include the ECL signal and the image of the membrane to visualize the molecular markers). For each pull-down experiment, the same samples were loaded on both nitrocellulose (that was stained with Ponceau) and PVDF membranes. - The top of the nitrocellulose membrane was first probed with Nup107, cut, and reprobed with anti-Nup133 His-Nup153 pull-down (nitro I or ii) Nup107 His-Nup153 pull-down (nitro I or ii) Nup133 (rehybr after Nup107) - The bottom of the nitrocellulose membrane was probed with anti-GAPDH antibody His-Nup153 pull-down (nitro i) GAPDH - The PVDF membrane was first cut and probed with Tpr, Nup96 and Nup85 His-Nup153 pull-down (PVDF i)Tpr-Nup96-Nup85 - The part of the PVDF membrane probed wiuth Nup98 was then reprobed with anti Nup98 His-Nup153 pull-down (PVDF i)_Nup98 (rehyb from Nup96)
Contributors: Rebecca Feltham, Kunzah Jamal, Tencho Tenev, Gianmaria Liccardi, Isabel Jaco, Celia Monteiro Domingues, Otto Morris, Sidonie Wicky John, Alessandro Annibaldi, Marcella Widya
... Original data of images and Western blot from the paper: Mind bomb regulates cell death during TNF signaling by suppressing RIPK1’s cytotoxic potential
PARN and TOE1 constitute a 3′ end maturation module for nuclear non-coding RNAs (A study of Son et al)
Contributors: Ahyeon Son, Jong-Eun Park, V. Narry Kim
... Original data of immunofluorescence, chemiluminescence, and autoradiography
Serine availability influences mitochondrial dynamics and function through lipid metabolism. Gao et al.
Contributors: Xia Gao, Jason Locasale
... All metabolomics and lipodomic data
Contributors: Lay Teng Ang
... Liver cells stained for protein expression by immunostaining in Figure 1G, 2H, 5B and 6D.
Asymmetric distribution of centromeric components in midgut stem cells and their role in cell types with varying differentiating potential
Contributors: Ana Garcia del Arco, Bruce A. Edgar, Sylvia Erhardt
... CENP-A is a histone variant critical for centromere function during mitosis. We have analyzed CENP-A in stem cells of the fly intestine and found an symmetric inheritance of pre-existing and newly synthesized CENP-A. CENP-A and its loading factor CAL1 are essential in differentiated cells, suggesting non-mitotic roles
Contributors: Yukiko Kitase, Julian Vallejo, Jianxun Yi, Harika Vemula
... We couldn't upload Dicom files for uCT analysis in Fig.2 due to the file size. Please contact corresponding authors if you are interested.
Contributors: Min-Dian Li
... Original data of images and Western blot. Cell Reports (2017), https://doi.org/10.1016/j.celrep.2017.09.051
Contributors: Yasuhiro Murakawa
... All the bed files used in a study by Yoshihara et al. entitled "Hotspots of de novo point mutations in induced pluripotent stem cells" in Cell Reports are provided. Please see the Experimental Procedure section and Table S2 and Table S3 for detail.
Contributors: Lior Levy, Leon Anavy, Roee Amit
... We use an oligonucleotide library of >10000 variants to identify an insulation mechanism encoded within a subset of σ54 promoters. Insulation manifests itself as reduced protein expression for a downstream gene that is expressed by transcriptional read-through. It is strongly associated with the presence of short CT-rich motifs (3-5 bp), positioned within 25 bp upstream of the Shine-Dalgarno (SD) motif of the silenced gene. We provide evidence that insulation is triggered by binding of the ribosome binding site (RBS) to the upstream CT-rich motif. We also show that in E.coli insulator sequences are preferentially encoded within σ54 promoters, suggesting an important regulatory role for these sequences in natural contexts. Our findings imply that sequence-specific regulatory effects that are sparsely encoded by short motifs may not be easily detected by lower throughput studies. Such sequence-specific phenomena can be uncovered with a focused OL design that mitigates sequence related variance, as exemplified herein.