Journal: bioRxiv

Article Title: Vascular endothelial growth factor-D improves lung vascular integrity during acute lung injury

doi: 10.1101/2024.12.16.628787

Figure Lengend Snippet: A) NICHES analysis of AFs, mesothelial cells, pericytes, gCap, and aCap in human microvascular niche yields a quantitative cell-cell signaling atlas of AF-aCap, AF-gCap, Mesothelial-aCap, Mesothelial-gCap, Pericyte-aCap, and Pericyte-gCap visualized by low-dimensional UMAP embedding. B) FeaturePlot of top differentially expressed L/R pairs ANGPT1 - TEK , SLIT2 - ROBO4 , VEGFD - KDR , BMP5-BMPR2 , ADM - RAMP2 , WNT5A - FZD6 , PTN - RACK1 , OXT - EDNRB , and ADCYAP1 - VIPR1 from NICHES analysis C) Top 10 differentially expressed L/R pairs between different cell-cell interaction signals processed through NICHES and plotted in DoHeatmap. D) Immunostaining of representative L/R pairs from AF to microvascular endothelial cells, including SLIT2 - ROBO4 , ADM - RAMP2 , BMP5 - BMPR2 , and VEGFD - VEGFR2 in human lung section. ITGA8 is a marker for AF; PRX is a marker for microvascular endothelial cells. Scale bar 20 μm.

Article Snippet: The protein levels of SLIT2, ADM, BMP5, and VEGFD in samples harvested from different timepoints were determined using corresponding ELISA kits: SLIT2 (# NB030384), ADM (# NBP2-78738), BMP5 (# NBP2-69996), VEGFD (# NBP2-78890), all from Novus Biologicals.

Techniques: Immunostaining, Marker

Journal: bioRxiv

Article Title: Vascular endothelial growth factor-D improves lung vascular integrity during acute lung injury

doi: 10.1101/2024.12.16.628787

Figure Lengend Snippet: A, B) The electrical resistance and permeability in HLMVECs after treatment with VEGF-D (1 μg/mL), adrenomedullin (ADM)(100 ng/mL), angiopoietin 1 (ANGPT1)(2 μg/mL), BMP-5 (1 μg/mL), SLIT2 (1 μg/mL), CXCL6 (1 μg/mL), Pleiotrophin (PTN)(1 μg/mL), SEMA6D (1 μg/mL), and TNF-α (1 ng/mL) were measured through electrical cell impendence sensing (ECIS) assay, and transwell assay, respectively. C) HLMVECs cultured on transwell treated with VEGF-D, followed by challenging with IL1β, TNF-α, or thrombin. The permeability was determined using a 20kDa FITC-dextran probe. D) Immunofluorescent images of F actin (red), and VE-Cadherin (green) in HLMVECs challenged with TNF-α or IL1β with or without VEGF-D. White boxes highlight VE-Cadherin junctions in all conditions. Scale bar 20 μm. E) Protein expression of phosphor-MLC in HLMVECs challenged with thrombin with different concentrations of VEGF-D was assessed through a western blot. Data presented as mean ± SEM. * and ** indicate p<0.05 and p<0.01, respectively.

Article Snippet: The protein levels of SLIT2, ADM, BMP5, and VEGFD in samples harvested from different timepoints were determined using corresponding ELISA kits: SLIT2 (# NB030384), ADM (# NBP2-78738), BMP5 (# NBP2-69996), VEGFD (# NBP2-78890), all from Novus Biologicals.

Techniques: Permeability, Transwell Assay, Cell Culture, Expressing, Western Blot

Journal: Journal of orthopaedic trauma

Article Title: The Effects of Episodic Alcohol Exposure on BMP2 Signaling During Tibia Fracture Healing

doi: 10.1097/BOT.0000000000001160

Figure Lengend Snippet: Illustration of selective components in the BMP signaling pathway, important for bone and cartilage formation and fracture healing. Bone morphogenetic proteins (BMP) induce intracellular signaling by binding the transmembrane receptors BMPR-II and BMPR-I. When these serine/threonine receptors are activated by BMPs, BMPR-I becomes phosphorylated. When both BMPR-II and BMPR-I are bound to BMPs and phosphorylated, downstream signaling is activated via the phosphorylation of transcription factors SMAD 1/5/8. Activated pSMAD 1/5/8 forms a complex with cofactor SMAD4, allowing for the regulated transcription of downstream target genes.

Article Snippet: BMP Signaling Assays Protein expressions of BMP2, BMPRII, BMPR1A, and the inhibitor Chordin were all assessed via the following commercially available ELISA kits: BMP2 (Boster Biological Technology, EKO312, Pleasanton, CA); BMPRII (Mybiosource, MBS701412, San Diego, CA); BMPRIA (Mybiosource, MBS720872); Chordin (Mybiosource, MBS2021328).

Techniques: Binding Assay, Phospho-proteomics

Journal: Experimental & Molecular Medicine

Article Title: IL-33/ST2 axis mediates hyperplasia of intrarenal urothelium in obstructive renal injury

doi: 10.1038/s12276-018-0047-8

Figure Lengend Snippet: a Total RNAs from UPJ tissues of control and obstructed kidneys were collected from C57BL/6 and IL33KO mice for qRT-PCR analyses ( n = 5 from each group). PC1 shows the log2 fold change in gene expression between UUO UPJ tissues and control (Ctrl) UPJ tissues from wild-type C57BL/6 mice. PC2 shows the log2 fold change in gene expression between IL33KO UUO UPJ tissues and C57BL/6 UUO UPJ tissues. The heatmap represents the expression of urothelial differentiation genes in UPJ tissues. b Western blot analysis of the SHH protein in UPJ tissues. c Immunofluorescent staining of SHH and IL-33 in UPJ tissues. d Levels of SHH, BMP4, and BMP5 in control and hydronephrotic urine samples were analyzed using ELISA ( n = 5 in each group). e Proposed working model for the mechanism by which the upregulation of IL-33 following obstructive renal injury induces type 2 immune responses and UPJ urothelium hyperplasia

Article Snippet: Mouse serum or urine levels of IL-33 (DY413, R&D System), IL-5 (DY405, R&D System), IL-13 (DY413, R&D System), SHH (DY461, R&D System), BMP4 (EK0316, Boster), and BMP5 (LS-F20454, LSBio) were analyzed using enzyme-linked immunosorbent assay (ELISA) kits according to the manufacturers’ instructions.

Techniques: Control, Quantitative RT-PCR, Gene Expression, Expressing, Western Blot, Staining, Enzyme-linked Immunosorbent Assay