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

Journal: Nature Communications

Article Title: Lysosome-mediated aggregation of galactose-deficient IgA1 with transferrin receptor 1 links to IgA nephropathy

doi: 10.1038/s41467-025-60819-w

Figure Lengend Snippet: a Confocal microscopy images of human mesangial cells stained with Proteostat dye. Aggregation was absent in cells in control group treated with PBS or HC-IgA1. Globular aggregation was presented in mesangial cells incubated with Gd-IgA1. The experiment was repeated three times independently with a similar result. b Gd-IgA1 was detected by anti-IgA1-AF488 antibody, and the colocalization with Proteostat-positive aggregated proteins with Gd-IgA1 was observed. c Lysosomal function assay was performed in mesangial cells treated with Cytochalasin D, HC-IgA1 and Gd-IgA1. d TFEB was found to be reduced in mesangial cells treated with Gd-IgA1, as shown by western blot analysis. Statistical analysis was performed using a two-sided Student’s t -test, n = 3 biological replicates; mean ± SD. e Western blot analysis revealed that Cathepsin B and Cathepsin D were decreased in mesangial cells treated with Gd-IgA1. Statistical analysis was performed using a two-sided Student’s t -test, n = 3 biological replicates; mean ± SD. f The medium of human mesangial cells stimulated with Gd-IgA1 and HC-IgA1 was tested with human cytokine array. The fluorescence of the chip in different treatments was demonstrated. g The fluorescent intensity of cytokines was quantified. Statistical analysis was performed using a two-sided Student’s t -test, n = 4 biological replicates; mean ± SD. h The medium of human mesangial cells stimulated with Gd-IgA1 and HC-IgA1 was tested with IL-6 ELISA. Statistical analysis was performed using a two-sided Student’s t -test, n = 3 biological replicates; mean ± SD. Source data are provided as a Source Data file.

Article Snippet: Mesangial cells were incubated with Gd-IgA1 or HC-IgA1 (1 mg/mL) for 24 h. Following this incubation period, the medium was replaced with fresh medium, and the cells were allowed to incubate for an additional 24 h. The conditioned medium was then collected and analyzed using a multiplex ELISA array that included a panel of 120 cytokines (Human Cytokine Array GS2000, RayBiotech Life, Inc.) following the manufacturer’s protocol.

Techniques: Confocal Microscopy, Staining, Control, Incubation, Functional Assay, Western Blot, Fluorescence, Enzyme-linked Immunosorbent Assay