psmad2  (Cell Signaling Technology Inc)

Journal: Research

Article Title: Mapping Immune-Inflammatory Niches on Zirconia Bone Implants: Single-Cell Transcriptomic Profiling

doi: 10.34133/research.1162

Figure Lengend Snippet: Ti and ZrO 2 implants induce early osseointegration and immune inflammation. (A) Transcriptional levels of osteogenesis-related and immune-inflammatory genes. Osteogenic genes: Runx2 , Osterix , and Tgfb1 . Immune-inflammatory genes: Il1b , Mmp9 , and Il23a . (B) Receiver operating characteristic (ROC) analysis based on bulk RNA-seq data. AUC, area under the curve. (C) Protein expression levels of osteogenesis-related and immune-inflammatory markers. Osteogenic proteins: RUNX2, OSX, TGFB1, pSMAD2/SMAD2, and pSMAD3/SMAD3. Immune-inflammatory proteins: IL1B, MMP9, and IL23A. (D) Statistical analysis of protein expression levels. Statistical significance was determined using one-way analysis of variance. RUNX2, RUNX family transcription factor 2; OSX, osterix; TGFB1, transforming growth factor beta 1; SMAD2, SMAD family member 2; SMAD3, SMAD family member 3; IL1B, interleukin 1 beta; MMP9, matrix metallopeptidase 9; IL23A, interleukin 23 subunit alpha.

Article Snippet: The membranes were blocked with 5% nonfat milk for 1 h, followed by overnight incubation at 4 °C with the following primary antibodies: RUNX2 (Bioss, bs-1134R, 1:1,000, China), OSX (Bioss, bs-25532R, 1:1,000, China), pSMAD2 (Bioss, bs-3419R, 1:1,000, China), pSMAD3 (Bioss, bs-3425R, 1:1,000, China), IL1B (Bioss, bs-0812R, 1:1,000, China), MMP9 (Bioss, bsm-54040R, 1:3,000, China), IL23A (Bioss, bs-1193R, 1:1,000, China), NOS2 (Bioss, bs-0162R, 1:2,000, China), CD206 (Bioss, bsm-55604R, 1:1,000, China), TGFB1 (Proteintech, 81746-2-RR, 1:1,000, China), SMAD2 (Proteintech, 12570-1-AP, 1:3,000, China), and SMAD3 (Proteintech, 30130-1-AP, 1:2,000, China).

Techniques: RNA Sequencing, Expressing

Journal: bioRxiv

Article Title: TGFβ determines epithelial tissue spacing by regulating mesenchymal condensation

doi: 10.64898/2026.03.16.712215

Figure Lengend Snippet: A. Schematic showing two possible mechanisms to spatially pattern proliferation in the embryonic lung. B. Representative immunoblots for pSMAD2 and GAPDH in lungs cultured with RepSox or vehicle control, and graph showing the normalized pSMAD2 band intensity from 3 replicates. Shown are the average and s.d. of normalized intensity of the pSMAD2 band. C. Fluorescence images of EdU incorporation in the epithelium and mesenchyme of lung explants cultured with RepSox or vehicle control. White arrows denote closely apposed branch stalks. Solid and dotted boxes denote the regions between b1 and b2, respectively. Scale bars, 100 µm. D. Graph showing the percentage of EdU-positive epithelial cells in lung explants cultured with RepSox or vehicle control. Shown are the average and s.d. of 6 lungs per condition. n.s. stands for not significant. *** p <0.001. E. Graph showing the percentage of EdU-positive mesenchymal cells in lung explants cultured with RepSox or vehicle control. Shown are the average and s.d. of 3 lungs per condition. ** p <0.005. F. Graph showing the density of mesenchymal cells near the tip in lung explants cultured with RepSox or vehicle control. Shown are the average and s.d. of at least 4 lungs per condition. * p <0.05.

Article Snippet: Samples were then incubated with primary antibodies against E-cadherin (1:100; BD Biosciences, 610182), pSMAD1/5/9 (1:100; Invitrogen, PA5-64711), pan-laminin (1:100; Invitrogen, PA1-16730), perlecan (1:100; Invitrogen, MA1-06821), or pSMAD2 (1:100; Cell Signaling, 18338).

Techniques: Western Blot, Cell Culture, Control, Fluorescence

Journal: bioRxiv

Article Title: TGFβ determines epithelial tissue spacing by regulating mesenchymal condensation

doi: 10.64898/2026.03.16.712215

Figure Lengend Snippet: ( A ) Fluorescence images of nuclei (blue) and EdU incorporation (red) in lung explants inserted with TGFβ-containing bead or vehicle control. The beads were inserted proximal to b1. In contrast to control, few EdU-positive mesenchymal cells are observed in TGFβ-containing bead-inserted lung. White arrows denote proximal stalk of b1. ( B ) Fluorescence images of epithelium in branch-implanted lung explants treated with RepSox or vehicle control. The distance between implanted and native branches is small in RepSox-treated lungs. Scale bars, 100 µm and 10 µm (inset). ( C ) Fluorescence images of nuclei in lung explants cultured for 12- or 24-hours after implantation of beads with different TGFβ concentrations. ( D ) Graph showing the sizes of condensations in bead-implanted lung explants with different initial TGFβ concentration and culture time. ( E ) Fluorescence images of nuclei (blue) and pSMAD2 (red) counterstained for E-cadherin (green). Strong circular pSMAD2 staining is observed near TGFβ-containing beads. ( F ) Fluorescence images of nuclei (blue) and perlecan (red) in lung explants inserted with TGFβ-containing bead or vehicle control. Strong perlecan staining is observed in mesenchymal condensations near TGFβ-containing beads. ( G ) Fluorescence images of nuclei in lung explants cultured with or without RepSox after being inserted with TGFβ-containing beads or vehicle control. Mesenchymal condensations are only observed in lungs inserted with TGFβ-containing beads and cultured without RepSox. ( H ) pSMAD2 staining and nuclear-masked pSMAD2 staining intensity near the branch. Scale bar, 50 µm. Subepithelial mesenchymal cells exhibit strong pSMAD2 staining. Cyan dotted lines denote condensations ( D, F, G ). Scale bars, 100 µm ( A, C, E, F, G ).

Article Snippet: Samples were then incubated with primary antibodies against E-cadherin (1:100; BD Biosciences, 610182), pSMAD1/5/9 (1:100; Invitrogen, PA5-64711), pan-laminin (1:100; Invitrogen, PA1-16730), perlecan (1:100; Invitrogen, MA1-06821), or pSMAD2 (1:100; Cell Signaling, 18338).

Techniques: Fluorescence, Control, Cell Culture, Concentration Assay, Staining