Journal: Endocrinology

Article Title: Msx Homeobox Genes Act Downstream of BMP2 to Regulate Endometrial Decidualization in Mice and in Humans

doi: 10.1210/en.2019-00131

Figure Lengend Snippet: Msx2 is a downstream target of BMP2 signaling in the uterus during decidualization. (A) The primary cultures of mouse endometrial stromal cells (MESCs) were transduced with adenovirus expressing GFP or BMP2. The cells were lysed at different time points, as indicated. Total RNA was isolated, and real-time PCR was performed to analyze the levels of Msx2. The relative levels of gene expression were determined by setting the expression level of the GFP-treated sample at 24 h to 1.0 (n = 3). Rplp0, encoding a ribosomal protein, was used to normalize the level of RNA. *P < 0.05. (B) The nucleotide positions of the SBEs on the Msx2 promoter were analyzed by ChIP. (C) Mouse stromal cells were treated with E + P or E, P, and BMP2 (E + P + BMP2) for 90 min. ChIP, using the Smad4 antibody, was performed, as described in “Materials and Methods.” Chromatin enrichment was quantified by real-time PCR using primers flanking the potential SBE in the Msx2 promoter and also a negative control region in the ORF of Msx2. Enrichments were normalized to 1% of input DNA. The experiment was repeated twice, and representative data are shown.

Article Snippet: The next day, the cells were either treated with E + P or E + P + BMP2 (recombinant human BMP2; R&D Systems; 355-BM-010) for 90 minutes.

Techniques: Transduction, Expressing, Isolation, Real-time Polymerase Chain Reaction, Gene Expression, Negative Control

Journal: Endocrinology

Article Title: Msx Homeobox Genes Act Downstream of BMP2 to Regulate Endometrial Decidualization in Mice and in Humans

doi: 10.1210/en.2019-00131

Figure Lengend Snippet: MSX1 and MSX2 mediate BMP2-induced HESC decidualization. The primary cultures of HESCs were established as described in “Materials and Methods.” The cells were transduced with adenovirus expressing GFP or BMP2. The cells were lysed at different time points as indicated. Total RNA was isolated, and real-time PCR was performed to analyze the levels of MSX1 and MSX2. The relative levels of gene expression were determined by setting the expression level on day 0 of the GFP-treated sample at 1.0. RPLP0, encoding a ribosomal protein, was used to normalize the level of RNA. Data were collected from three independent clinical samples, which were subjected to the same experimental conditions. *P < 0.05; **P < 0.005.

Article Snippet: The next day, the cells were either treated with E + P or E + P + BMP2 (recombinant human BMP2; R&D Systems; 355-BM-010) for 90 minutes.

Techniques: Transduction, Expressing, Isolation, Real-time Polymerase Chain Reaction, Gene Expression

Journal: Immunity

Article Title: Stellate Cells, Hepatocytes, and Endothelial Cells Imprint the Kupffer Cell Identity on Monocytes Colonizing the Liver Macrophage Niche

doi: 10.1016/j.immuni.2019.08.017

Figure Lengend Snippet:

Article Snippet: Recombinant Human/Mouse/Rat BMP-2 Protein , RnD Systems , Cat#355-BM-010.

Techniques: Control, Recombinant, Blocking Assay, Irradiation, Enzyme-linked Immunosorbent Assay, cDNA Synthesis, Microarray, Software, Microscopy

Journal: Molecular cell

Article Title: An Epstein-Barr virus protein interaction map reveals NLRP3 inflammasome evasion via MAVS UFMylation

doi: 10.1016/j.molcel.2023.05.018

Figure Lengend Snippet: (A) Immunoblot of ASC oligomerization from P3HR-1 expressing the indicated sgRNA, uninduced or 4HT-induced for 24 h. (B) Immunoblot analysis of WCL from P3HR-1 expressing the indicated sgRNA and 4HT-induced, as indicated. (C) Mean ± SEM caspase-1 activity normalized by live cell number from n = 3 replicates of P3HR-1 expressing the indicated sgRNA and 4HT-treated for 24 h, as indicated. (D) Immunoblot of 1% input and anti-FLAG-MAVS complexes from 293T co-transfected with FLAG-MAVS, BILF1, or BXLF1 cDNAs for 24 h, as indicated. (E) PEAKS software identification of potential MAVS post-translational modification sites. Putative UFMylation sites were identified at lysines 362, 371, and 461. MAVS residues 321–500 are depicted. Black vertical lines represent individual peptide sequencing events. (F) Immunoblot of anti-EGFP-MAVS immunopurified from 293T co-transfected with the indicated MAVS, BILF1, and UFM1 cDNAs for 24 h. (G) Immunofluorescence analysis of wild type or K461R MAVS subcellular localization in 293T co-transfected with MAVS and BILF1 cDNAs for 24 h. (H) Immunoblot of WCL from 293T expressing MAVS and BILF1 cDNA, nigericin stimulated for 24 h, as indicated. (I) Immunoblot of 1% input and anti-HA immunopurified EGFP-MAVS and UFL1 from 293T transfected with BILF1 and EGFP-MAVS cDNAs for 24 h, as indicated. (J) AlphaFold multimer model highlighting the predicted BILF1 and MAVS interaction domain and residues. (K) Immunoblot of WCL from 293T expressing wild type, N- (ΔN) or C- (ΔC) terminal tail deletion mutant BILF1, nigericin stimulated for 24 h, as indicated. (L) Schematic of BILF1 NLRP3 inflammasome inhibition. Student’s t test was performed, with ***p < 0.001. *p < 0.05. See also and . Immunoblots are representative of n = 2.

Article Snippet: Anti-UFM1 rabbit polyclonal antibody , Proteintech , cat# 15883-1-AP; RRID: AB_2878195.

Techniques: Western Blot, Expressing, Activity Assay, Transfection, Software, Modification, Sequencing, Immunofluorescence, Mutagenesis, Inhibition

Journal: Molecular cell

Article Title: An Epstein-Barr virus protein interaction map reveals NLRP3 inflammasome evasion via MAVS UFMylation

doi: 10.1016/j.molcel.2023.05.018

Figure Lengend Snippet:

Article Snippet: Anti-UFM1 rabbit polyclonal antibody , Proteintech , cat# 15883-1-AP; RRID: AB_2878195.

Techniques: Recombinant, Magnetic Beads, Transfection, Sequencing, Modification, Mass Spectrometry, Produced, Protease Inhibitor, Selection, Purification, Gel Extraction, SYBR Green Assay, Reporter Assay, Protein Quantitation, CRISPR, Amplification, Molecular Cloning, Plasmid Preparation, Software