Journal: RNA biology

Article Title: Nucleolar disruption leads to the spatial separation of key 18S rRNA processing factors.

doi: 10.4161/rna.18811

Figure Lengend Snippet: Figure 3. Localization of SSU processome proteins after treatment with ActD. (A) HeLa cells, either control or treated with ActD (indicated at the top) were fixed and then analyzed by immunofluorescence using anti-MPP10, -BMS1, -UTP13 and -tUTP10 antibodies. Each set of cells was also incubated with mouse monoclonal anti-fibrillarin antibodies. (B) HeLa cells were treated with ActD, fixed and then analyzed using anti-fibrillarin and anti-UTP12 antibodies (left set of panels). HeLa cells, expressing GFP-fibrillarin, were treated with ActD and then analyzed using anti-RPA135 antibodies (right set of panels). Note that the localization of UTP12 and RPA135 in untreated cells was basically the same as that of UTP13 and tUTP10. (C) HEK293 cells expressing the FLAG-tagged proteins (as indicated) were fixed and then analyzed by immunofluorescence using anti-FLAG and anti-fibrillarin antibodies. The identity of the protein detected is indicated in each panel. A color overlay of the two images, with DAPI staining (blue), is shown at the bottom (merge). Bar: 1 mm.

Article Snippet: Antibodies raised against hU3–55K, MPP10, UTP12, BMS1, UTP13 and tUTP10 were described previously.16,22 Anti-RPA135 and anti-nucleolin antibodies were purchased from Santa Cruz biotechnology and Abcam, respectively.

Techniques: Control, Immunofluorescence, Incubation, Expressing, Staining

Journal: Bioactive Materials

Article Title: Synergistic enhancement of immunological responses triggered by hyperthermia sensitive Pt NPs via NIR laser to inhibit cancer relapse and metastasis

doi: 10.1016/j.bioactmat.2021.05.030

Figure Lengend Snippet: Schematic illustration of Pt NPs conjugated with BMS-1 through hyperthermia-sensitive linkage for NIR-controlled release of inhibitor and exposure of Mal . A thermal-sensitive release and Mal deprotection procedure is achieved by the Retro D-A reaction. Exposed Mal on the surface of Pt NPs captures the antigens from ablated tumor cells and promotes antigen presentation. The released BMS-1 alleviates T cell exhaustion and induces infiltration of effector T cells.

Article Snippet: Maleimide-PEG 2k -COOH (Mal-PEG 2k -COOH) and PD-L1 inhibitor (BMS-1) were purchased from Xi'an ruixi Biological Technology Co., Ltd, China. mPEG 2k -OH was purchased from Sigma-Aldrich.

Techniques:

Journal: PLoS Genetics

Article Title: BMS1 Is Mutated in Aplasia Cutis Congenita

doi: 10.1371/journal.pgen.1003573

Figure Lengend Snippet: a. Representative images of individuals with ACC in a five-generation pedigree with autosomal dominant inheritance pattern and full penetrance. (Left image: 1 year old with hypertrophic scar at site of congenital ACC of the scalp; Middle image: father of 1 year old of left image with scar at site of congenital ACC; Right image: daughter of this father on day 1 after birth with localized erosion on vertex of scalp (indicated with arrow in pedigree; this member was born after the linkage study was completed and carried the disease allele as well)). Family members' DNA used for genome-wide SNP genotyping indicted with black stars; additional family members' DNA included for confirmation of linkage with microsatellite markers indicated in red stars. b. Histogram of multi-point LOD scores along chromosome 10 shows a single chromosomal region with LOD scores >2 on chromosome 10q11. c. A heterozygous missense mutation resulting in a G>A nucleotide change in the BMS1 gene was identified in all affected members with ACC in this family (c.2789G>A). d. The c.2789G>A mutation results in a Arg-to-His amino acid change (p.R930H) of a conserved Arg residue in BMS1. e. The p.R930H amino acid change affects an Arg residue within the putative C-terminal GAP domain of BMS1. Representative image of the domains of BMS1 with its GTPase domain at the N-terminus (adapted from ). f. Murine embryonic sections showing the site of skin formation at the vertex area of the scalp at E13.5 (f. and g.). BMS1 expression is seen in all cells, including the proliferative epidermis (C-terminus of BMS1 in red, pH3b green). C-term indicates labeling with the monoclonal anti-BMS1 antibody that recognizes the C-terminus of BMS1. pH3b indicates staining for the proliferation marker phospho-Histone 3B (Ser10) (pH3b, green). g. Co-localization of BMS1 C-terminal (C-term red) and N-terminal domains (N-term green) at E13.5 of embryonic murine scalp at E13.5. Nuclei are labeled with DAPI. All images are acquired with a 20× objective.

Article Snippet: Pretested SRSF3 and BMS1 qPCR primers were used from SABiosciences (Qiagen) and results were normalized to 36B4 transcript levels.

Techniques: Genome Wide, Mutagenesis, Residue, Expressing, Labeling, Staining, Marker

Journal: PLoS Genetics

Article Title: BMS1 Is Mutated in Aplasia Cutis Congenita

doi: 10.1371/journal.pgen.1003573

Figure Lengend Snippet: a. Pre-rRNA processing pathways in human cells (adapted from ). Two pathways coexist depending on the order of cleavage in the 5′-ETS (sites A 0 and A 1 ) and ITS1 (A 2 ). Nomenclature of the cleavage sites follows Hadjiolova et al. (1993) and Rouquette et al. (2005) , . b. Pulse-chase labeling of pre-rRNAs with L-[ methyl - 3 H]methionine shows that ACC BMS1(p.R930H) fibroblasts (ACC) form all pre-rRNA species compared to control fibroblasts (WT), but retain increased 45S and 30S pre-rRNAs (seen at 180 minutes after pulse-labeling). Relative values for bands normalized to 28S rRNA band are shown (fold-change compared to WT). c. Magnification of 45S pre-rRNAs 180 minutes after pulse labeling shows retained 45S pre-rRNA in ACC BMS1(p.R930H) fibroblasts compared to control fibroblasts (WT). d. Northern blot analysis of pre-rRNA processing using a radioactive ITS-1 probe. RNA from ACC fibroblasts was used carrying the BMS1 p.R930H mutation, as well as from control fibroblasts (WT). The Northern blot shown on the left side for the WT and ACC cells was exposed overnight, while the image on the right side shows the same blot after a 4 hour exposure. In addition, RNA from fibroblasts stably transfected with an inducible BMS1 shRNA vector was used, after shRNA-mediated knockdown of BMS1 transcripts was induced by doxycycline (doxy) treatment. Ethidiumbromide stained gels prior to blotting (bottom) confirm equal loading of RNA. Quantitation of band intensity ratios expressed as relative values (fold-change compared to WT or untreated cells). e. Doxycyline-inducible knockdown of BMS1 by shRNA lentivirus (shBMS1). Doxycycline treatment for 48 hours results in ∼60% reduction of BMS1 transcript levels. * P-value<0.05.

Article Snippet: Pretested SRSF3 and BMS1 qPCR primers were used from SABiosciences (Qiagen) and results were normalized to 36B4 transcript levels.

Techniques: Pulse Chase, Labeling, Control, Northern Blot, Mutagenesis, Stable Transfection, Transfection, shRNA, Plasmid Preparation, Knockdown, Staining, Quantitation Assay