Journal: Scientific Reports

Article Title: SETD4-expressing cells contribute to pancreatic development and response to cerulein induced pancreatitis injury

doi: 10.1038/s41598-021-92075-5

Figure Lengend Snippet: Identification of SETD4 + cells in embryonic SETD4-CreER T2 ; Rosa26 mTmG/ + mice ( a ) and lineage tracing of SETD4 + cells in SETD4-Cre ; Rosa26 mTmG/ + mice ( b – f ). ( a ) Representative immunofluorescence and quantification for recombinant (SETD4 + ) cells 24 h after tamoxifen-induction of at embryo day 13.5 (E13.5), E16.5 and E19.5 mouse pancreases. ( b ) Representative wholemounts fluorescent images of pancreases in E15.5, P0 and P56 mice. Scale bar for each image, 0.5, 1, 4 mm, respectively. The white arrow indicates the pancreas. ( c ) Representative immunofluorescence and quantification of GFP + recombinant cells in the total (DAPI + ) cells of E15.5, P0 and P56 pancreas. Scale bar for each image, 10 μm. ( d ) Representative immunofluorescence for SETD4 + cells descendants (recombinant cells) with Sox9 and Pdx1 in E9.0 pancreatic bud. Scale bar for each image, 50 μm. ( e ) Representative immunofluorescence for SETD4 + cells descendants (recombinant cells) with Sox9, Nkx6.1, Cpa1 and Pdx1 in the E15.5 pancreas. Scale bar for each image, 50 μm. ( f ) Representative immunofluorescence for recombinant cells with amylase in the acinar, CK19 in the duct and insulin, glucagon, somatostatin and ghrelin in the islet in P56 pancreas. All data are represented as mean ± SD. n = 3 mice. Nuclei were stained with DAPI.

Article Snippet: Slides were mounted in Antifade Mounting Medium with DAPI (Beyotime P0131).

Techniques: Immunofluorescence, Recombinant, Staining

Journal: Scientific Reports

Article Title: SETD4-expressing cells contribute to pancreatic development and response to cerulein induced pancreatitis injury

doi: 10.1038/s41598-021-92075-5

Figure Lengend Snippet: Lineage tracing of SETD4 + cells in P3 and 8 weeks SETD4-CreER T2 ; Rosa26 mTmG/ + mice. ( a ) Schedule and lineage tracing of SETD4 + cells in P3 SETD4-CreER T2 ; Rosa26 mTmG/ + mice. Neonatal mice were treated with 2 consecutive doses of TAM and then analyzed at 2 time points: 24 h or 8 weeks post-induction with TAM. Representative immunofluorescence and quantification of single (1), doublet (2), and clones of multi-cells (≥ 3) at the respective time points are shown. Solid arrows: single cell (1), hollow arrows: multi-cell (≥ 3) clones. Scale bar in the lineage, 80 μm. ( b ) Quantification for Ki67 and PCNA of GFP + recombinant cells in the acinar, duct or islet compartments from adult pancreases. ( c ) Schedule of lineage tracing of SETD4 + cells in 8 weeks SETD4-CreER T2 ; Rosa26 mTmG/ + mice. Adult mice were treated with 2 consecutive doses of TAM and then analyzed at 4 time points: 24 h, 1, 4 and 10 months post-induction with TAM. ( d ) Representative immunofluorescence for amylase, CK19, insulin, glucagon, somatostatin and ghrelin in pancreas of adult (8 weeks) mice pancreases at 4 months after TAM-induction. Scale bar for all merged images, 15 μm. Scale bar for all spilt images, 10 μm. ( e ) Quantification of recombinant GFP + cells of total (DAPI + ) pancreatic cells in adult (P56) mice 1 month and 4 months after TAM-induction. ( f ) Representative immunofluorescence of lineage tracing of SETD4 + cells 10 months after TAM-induction in adult SETD4-CreER T2 ; Rosa26 mTmG/ + mice. Solid arrows: single cell (1), hollow arrows: multi-cell (≥ 3) clones. Scale bar in the lineage, 40 μm. ( g ) Isolation by fluorescence activated cell sorting (FACS) of GFP + and GFP - cells from SETD4-CreER T2 ; Rosa26 mTmG/ + mice at 24 h after TAM-induction. GFP + and GFP - cells in duct compartment were sorted and then cultured in 3-Dimentional Matrigel. Representative formation of cell-organized organoids by SETD4 + cells (green) in a 3-Dimentional Matrigel-based culture over a month. Scale bar for each image, 15 μm. All data are represented as mean ± SD. * p < 0.05. *** p < 0.001. ns not significant. n = 4 mice. Arrows indicate recombinant GFP + cells (green). Nuclei were stained with DAPI.

Article Snippet: Slides were mounted in Antifade Mounting Medium with DAPI (Beyotime P0131).

Techniques: Immunofluorescence, Clone Assay, Recombinant, Isolation, Fluorescence, FACS, Cell Culture, Staining

Journal: International Journal of Molecular Sciences

Article Title: Acceleration of TAA-Induced Liver Fibrosis by Stress Exposure Is Associated with Upregulation of Nerve Growth Factor and Glycopattern Deviations

doi: 10.3390/ijms22105055

Figure Lengend Snippet: ( A ) For an analysis of the glycophenotype, lectins were used to detect different types of glycosylation. O-glycan structures were recognized by Helix pomatia agglutinin (HPA; Tn-antigen) and Arachis hypogaea lectin (PNA; core 1). Lycopersicon esculentum lectin (LEA) recognizes polyLAcNac sequences. Phaseolus vulgaris lectin (PHA-L) recognizes β1-6GlcNAc-branched complex N-glycans and Phaseolus vulgaris lectin E subunit (PHA-E) identifies glycans with a bisecting modification. Fucosylation was determined by Lotus tetragonolobus lectin (LTL) and finally sialyation was determined using the Maackia amurensis lectin (MAA) and Sambucus nigra agglutinin (SNA-I) which bind to α2,3- and α2,6-linked sialic acid, respectively. Quantification of O-glycan ( B ), fucosylation ( C ), N-glycosylation ( D ), or sialylated glycan ( E ) was performed on liver sections of mice with fibrosis (TAA), fibrosis and stress (TAA + stress), and fibrosis and NGF administration (TAA + NGF). Mean fluorescence intensity (MFI) ± S.E.M is shown. * p < 0.05 and *** p < 0.001 vs. TAA (Kruskal–Wallis test). Expression of MGAT5 ( F ) and C1GALT1 ( G ) in patients with fibrosis due to HCV (GSE6764), HBV (GSE84044), or NAFLD (GSE49541). * p < 0.05 and ** p < 0.01 (Mann–Whitney test).

Article Snippet: Quantitative analysis of collagen content was performed by computerized morphometric analysis on samples stained with Sirius red and digitally scanned by a high-resolution bright field and fluorescence slide scanner (Pannoramic MIDI BF/FL, 3DHISTECH Ltd., Budapest, Hungary).

Techniques: Glycoproteomics, Modification, Fluorescence, Expressing, MANN-WHITNEY