Journal: Current pharmaceutical biotechnology

Article Title: Oral fluid vs. Urine Analysis to Monitor Synthetic Cannabinoids and Classic Drugs Recent Exposure

doi: 10.2174/1389201018666171122113934

Figure Lengend Snippet: Total ion chromatogram of 11 synthetic cannabinoids and 13 metabolites in urine sample at 15 ng/mL. 1, AB-PINACA 5-Pentanoic acid; 2, A-796260; 3, AB-FUBINACA; 4, JWH-250 4-Hydroxypentyl; 5, JWH-073 4-Butanoic acid; 6, AB-PINACA; 7, JWH-018 5-Pentanoic acid; 8, AM2201 4-Hydroxypentyl; 9, MAM2201 4-Hydroxypentyl; 10, JWH-019 5-Hydroxyhexyl; 11, UR-144 5-Pentanoic acid; 12, JWH-122 4-Hydroxypentyl; 13, XLR-11 4-Hydroxypentyl; 14, UR-144 4-Hydroxypentyl; 15, 5-Fluoro PB-22; 16, JWH-210 4-Hydroxypentyl; 17, APINACA (AKB-48) 5-Hydroxypentyl; 18, PB-22; 19, MAM2201; 20, JWH-073; 21, XLR-11; 22, JWH-018; 23, UR-144; 24, APINACA (AKB-48).

Article Snippet: Synthetic cannabinoids (A-796260, AB-FUBINACA, AB-PINACA, APINACA (AKB-48), JWH-018, JWH-073, MAM2201, PB-22, 5-Fluoro PB-22, UR-144, XLR-11) and metabolites standards (AB-PINACA 5-Pentanoic acid, APINACA (AKB-48) 5-Hydroxypentyl, AM2201 4-Hydroxypentyl, JWH-018 5-Pentanoic acid, JWH-019 5-Hydroxyhexyl, JWH-073 4-Butanoic acid, JWH-122 4-Hydroxypentyl, JWH-210 4-Hydroxypentyl, JWH-250 4-Hydroxypentyl, MAM2201 4-Hydroxypentyl, UR-144 4-Hydroxypentyl, UR-144 5-Pentanoic acid, XLR-11 4-Hydroxypentyl) and internal standards (IS) (JWH-250 4-Hydroxypentyl-d 5 , AM2201 4-Hydroxypentyl-d 5 , and JWH-210 4-Hydroxypentyl-d 5 ) were purchased from Cerilliant (Round Rock, TX) at 100 μg/mL in methanol or acetonitrile.

Techniques:

Journal: Current pharmaceutical biotechnology

Article Title: Oral fluid vs. Urine Analysis to Monitor Synthetic Cannabinoids and Classic Drugs Recent Exposure

doi: 10.2174/1389201018666171122113934

Figure Lengend Snippet: Total ion chromatogram of 11 synthetic cannabinoids in oral fluid sample at 2.5 ng/mL. 1, A-796260; 2, AB-FUBINACA; 3, AB-PINACA; 4, 5-Fluoro PB-22; 5, PB-22; 6, MAM2201; 7, JWH-073; 8, XLR-11; 9, JWH-018; 10, UR-144; 11, APINACA (AKB-48).

Article Snippet: Synthetic cannabinoids (A-796260, AB-FUBINACA, AB-PINACA, APINACA (AKB-48), JWH-018, JWH-073, MAM2201, PB-22, 5-Fluoro PB-22, UR-144, XLR-11) and metabolites standards (AB-PINACA 5-Pentanoic acid, APINACA (AKB-48) 5-Hydroxypentyl, AM2201 4-Hydroxypentyl, JWH-018 5-Pentanoic acid, JWH-019 5-Hydroxyhexyl, JWH-073 4-Butanoic acid, JWH-122 4-Hydroxypentyl, JWH-210 4-Hydroxypentyl, JWH-250 4-Hydroxypentyl, MAM2201 4-Hydroxypentyl, UR-144 4-Hydroxypentyl, UR-144 5-Pentanoic acid, XLR-11 4-Hydroxypentyl) and internal standards (IS) (JWH-250 4-Hydroxypentyl-d 5 , AM2201 4-Hydroxypentyl-d 5 , and JWH-210 4-Hydroxypentyl-d 5 ) were purchased from Cerilliant (Round Rock, TX) at 100 μg/mL in methanol or acetonitrile.

Techniques:

Journal: Current pharmaceutical biotechnology

Article Title: Oral fluid vs. Urine Analysis to Monitor Synthetic Cannabinoids and Classic Drugs Recent Exposure

doi: 10.2174/1389201018666171122113934

Figure Lengend Snippet: Urine samples (n=70) positive results for cannabis, cocaine, opiates, benzodiazepines, amphetamines and synthetic cannabinoids (AB-FUBINACA, PB-22, 5-Fluoro-PB-22, UR-144 metabolites). Cutoffs in urine were 1 ng/mL for synthetic cannabinoids, 50 ng/mL for cannabis, 200 ng/mL for benzodiazepines and 300 ng/mL for cocaine, opiates and amphetamines.

Article Snippet: Synthetic cannabinoids (A-796260, AB-FUBINACA, AB-PINACA, APINACA (AKB-48), JWH-018, JWH-073, MAM2201, PB-22, 5-Fluoro PB-22, UR-144, XLR-11) and metabolites standards (AB-PINACA 5-Pentanoic acid, APINACA (AKB-48) 5-Hydroxypentyl, AM2201 4-Hydroxypentyl, JWH-018 5-Pentanoic acid, JWH-019 5-Hydroxyhexyl, JWH-073 4-Butanoic acid, JWH-122 4-Hydroxypentyl, JWH-210 4-Hydroxypentyl, JWH-250 4-Hydroxypentyl, MAM2201 4-Hydroxypentyl, UR-144 4-Hydroxypentyl, UR-144 5-Pentanoic acid, XLR-11 4-Hydroxypentyl) and internal standards (IS) (JWH-250 4-Hydroxypentyl-d 5 , AM2201 4-Hydroxypentyl-d 5 , and JWH-210 4-Hydroxypentyl-d 5 ) were purchased from Cerilliant (Round Rock, TX) at 100 μg/mL in methanol or acetonitrile.

Techniques:

Journal: Frontiers in Endocrinology

Article Title: VHL-P138R and VHL-L163R Novel Variants: Mechanisms of VHL Pathogenicity Involving HIF-Dependent and HIF-Independent Actions

doi: 10.3389/fendo.2022.854365

Figure Lengend Snippet: Differences in mRNA fibronectin expression for P138R and L163R pVHL variants with similar disrupted deposition patterns. (A) Fibronectin mRNA expression of 786-O, WT, P138R, and L163R cell lines. Results are presented as fold change compared to WT cells. Values are expressed as ± SD of three independent experiments performed in triplicate. ns, not significant; *p = 0.0011, **p = 0.0030, one-way ANOVA and Tukey’s posttest. (B) Cell lines were cultured on coverslips to assess fibronectin deposition with anti-fibronectin Cy5 conjugated (in red) by immunofluorescence. Nuclei were dyed with 5 μg/ml Hoechst as shown in blue. Images were taken at ×40 on a Carl-Zeiss AxioScope A1 microscope.

Article Snippet: Nuclei were dyed with Hoechst (5 µg/ml), and pictures were taken on a Carl-Zeiss AxioScope A1 microscope.

Techniques: Expressing, Cell Culture, Immunofluorescence, Microscopy

Journal: Life Science Alliance

Article Title: Dimerization of kringle 1 domain from hepatocyte growth factor/scatter factor provides a potent MET receptor agonist

doi: 10.26508/lsa.202201424

Figure Lengend Snippet: (A) Possible receptor-binding modes and stoichiometries for hepatocyte growth factor/scatter factor and NK1 in the presence of heparin. (B) Schematic representation of hepatocyte growth factor/scatter factor, NK1, K1K1, K1K1 variants, K1H6, full-length c-MET, and the MET567 fragment. Individual domains (boxes) with positions of domain boundaries indicated above. CR, cysteine rich; Ig, immunoglobulin-like; SPH, serine protease homology; TK, tyrosine kinase. The transmembrane domain is indicated in red.

Article Snippet: For dose–response analysis, 8-wk-old FVB mice weighing 19–21 g (Charles River) were i.p. injected with 0.1, 0.5, 1, and 5 μg of K1K1 diluted in 100 μl of PBS and euthanized per cervical dislocation 10 min after injection.

Techniques: Binding Assay

Journal: Life Science Alliance

Article Title: Dimerization of kringle 1 domain from hepatocyte growth factor/scatter factor provides a potent MET receptor agonist

doi: 10.26508/lsa.202201424

Figure Lengend Snippet: (A) Amino acid sequence of K1K1H6 with SEVE linker in dark grey and poly-histidine tag in green. (B) The crystal structure of the two molecules of K1K1 (cyan) and K1K1H6 (green) showing the straight conformation and the N-terminus (NH 2 ) and C-terminus (COOH) located centrally in the linker region and a nearly identical overall structure with a root mean square deviation ranging from 0.8 to 1.8 Å. The C-terminal poly-histidine tag of K1K1H6 is making contacts with residues in the N-terminal kringle domain. (C) Surface representations of single kringle domains of K1K1 showing the location of the residues involved in MET binding, as defined by 40, shown in blue. The more lateral position of the heparin-binding site is shown in orange with the residues targeted by reverse-charge mutation in K1K1S2 (K10E, R12E) and K1K1S4 (K10E, R12E, K48E, R59E) indicated.

Article Snippet: For dose–response analysis, 8-wk-old FVB mice weighing 19–21 g (Charles River) were i.p. injected with 0.1, 0.5, 1, and 5 μg of K1K1 diluted in 100 μl of PBS and euthanized per cervical dislocation 10 min after injection.

Techniques: Sequencing, Binding Assay, Mutagenesis

Journal: Life Science Alliance

Article Title: Dimerization of kringle 1 domain from hepatocyte growth factor/scatter factor provides a potent MET receptor agonist

doi: 10.26508/lsa.202201424

Figure Lengend Snippet: (A) Chromatography profile of the elution of K1K1 from the Heparin HiTrap column showing the 280 nm absorbance in blue and the gradient of eluent (1 M NaCl) in green. (B) UPLC/MS analysis of K1K1 after size exclusion chromatography showing estimated molecular mass nearly identical to the mass as predicted based on the amino acid sequence (19195.24 versus 19,207.69 D). (C) UPLC/MS analysis of K1K1H6 after size exclusion chromatography and showing estimated molecular mass nearly identical to the mass as predicted based on the amino acid sequence (20,018.24 versus 20,030.53 D). (D) Coomassie-stained SDS–PAGE gel showing the different recombinant proteins used in this study (see design in ). About 5 μg of protein was loaded in reducing sample buffer in each lane. Source data are available for this figure.

Article Snippet: For dose–response analysis, 8-wk-old FVB mice weighing 19–21 g (Charles River) were i.p. injected with 0.1, 0.5, 1, and 5 μg of K1K1 diluted in 100 μl of PBS and euthanized per cervical dislocation 10 min after injection.

Techniques: Chromatography, Size-exclusion Chromatography, Sequencing, Staining, SDS Page, Recombinant

Journal: Life Science Alliance

Article Title: Dimerization of kringle 1 domain from hepatocyte growth factor/scatter factor provides a potent MET receptor agonist

doi: 10.26508/lsa.202201424

Figure Lengend Snippet: (A) Semi-transparent surface presentation of K1K1 alignment based on one kringle domain with that of the kringle domain of one NK1 protomer. The alignment projects the second K1K1 kringle across and positions it close to but not in the same position as the second kringle domain in the NK1 dimer (blue). (B) Two detailed 90° views of the K1K1-NK1 kringle alignment showing the kringle of K1K1 in pink and the kringle of the NK1 protomer in magenta. The straight, stretched-out conformation of K1K1 misaligns the second kringle domain by a rotation of 109.7°, leading to a translation of 13.9 Å. (C) The linker based on the naturally occurring linker sequence between kringle 1 and kringle 2 in hepatocyte growth factor/scatter factor, SEVE, is straight in K1K1 (pink) and has a different conformation in two NK2 structures available ( 3HN4 in yellow and 3SP8 in green). Numbering is given in italic from the last cysteine of kringle 1 (C206 in NK2) until the first cysteine in kringle 2 (C211 in NK2) with equivalent K1K1 residues in bold. (D) Superimposition of K1K1 SAXS envelop with the two kringle domains of the protomers in the NK1 dimer. (E) Additional side and top views of the K1K1 SAXS envelope (pink) placed within the cryo-EM map of the NK1 dimer (blue)-SEMA domain (grey) complex (7mob.pdb). Source data are available for this figure.

Article Snippet: For dose–response analysis, 8-wk-old FVB mice weighing 19–21 g (Charles River) were i.p. injected with 0.1, 0.5, 1, and 5 μg of K1K1 diluted in 100 μl of PBS and euthanized per cervical dislocation 10 min after injection.

Techniques: Sequencing, Cryo-EM Sample Prep

Journal: Life Science Alliance

Article Title: Dimerization of kringle 1 domain from hepatocyte growth factor/scatter factor provides a potent MET receptor agonist

doi: 10.26508/lsa.202201424

Figure Lengend Snippet: (A) The measured SAXS envelope is not compatible with the elongated K1K1 crystal structure (pink) but perfectly accommodates a bend conformation of K1K1 (cyan). To fit the SAXS envelope, the linker region is bent by roughly 60°. (B) The ab initio SAXS envelope of the 1:1 complex of K1K1, and MET567 receptor fragment shows a “pan-handle” extension, which accommodates K1K1. The CR domain is partially protruding from the bottom of the envelope, and extra volume might be occupied by glycosylated side chains not modelled. Images generated with UCSF Chimera. (C) The SAXS envelope of K1K1 in solution (pink) overlaps well with part of the cryo-EM map (emd_23923.map) accommodating the two kringle domains of the NK1 dimer (blue). Cryo-EM map for 7mob.pdb.

Article Snippet: For dose–response analysis, 8-wk-old FVB mice weighing 19–21 g (Charles River) were i.p. injected with 0.1, 0.5, 1, and 5 μg of K1K1 diluted in 100 μl of PBS and euthanized per cervical dislocation 10 min after injection.

Techniques: Generated, Cryo-EM Sample Prep

Journal: Life Science Alliance

Article Title: Dimerization of kringle 1 domain from hepatocyte growth factor/scatter factor provides a potent MET receptor agonist

doi: 10.26508/lsa.202201424

Figure Lengend Snippet: (A) Phosphorylation analysis by Western blot on Hela cell lysates after stimulation with ligands for 10 min at concentrations indicated above each lane. Loading controls are based on total MET, total Akt, and total ERK present in each lane. (B, C) AlphaScreen measurements of p-Akt and (C) p-ERK activation in HeLa cells after 10 min stimulation with K1K1, K1K1H6, HGF/SF, and K1H6. (D) Binding determination using the AlphaScreen saturation binding assay. Seven concentrations of K1K1H6 were tested on several different concentrations of MET-Fc (AA 25–922) . Shown is the binding of K1K1H6 to 3 nM MET-Fc. (E) Ligand induced MDCK survival after overnight treatment with the apoptotic inducer anisomycin. Indicated is the percentage of viable cells compared with no-anisomycin treatment after exposure to HGF/SF, K1K1, K1K1H6, and NK1 at different concentrations. Source data are available for this figure.

Article Snippet: For dose–response analysis, 8-wk-old FVB mice weighing 19–21 g (Charles River) were i.p. injected with 0.1, 0.5, 1, and 5 μg of K1K1 diluted in 100 μl of PBS and euthanized per cervical dislocation 10 min after injection.

Techniques: Phospho-proteomics, Western Blot, Amplified Luminescent Proximity Homogenous Assay, Activation Assay, Binding Assay, Saturation Assay

Journal: Life Science Alliance

Article Title: Dimerization of kringle 1 domain from hepatocyte growth factor/scatter factor provides a potent MET receptor agonist

doi: 10.26508/lsa.202201424

Figure Lengend Snippet: (A) AlphaScreen cross titration assay using MET-Fc chimera (0–10 nM) and K1K1H6 (0–300 nM) dilutions was performed using protein A–coated Alpha acceptor beads with Ni-NTA–coated Alpha donor beads. The Alpha signal is expressed in CPS (photon counts per seconds). Measurements are expressed as technical duplicates (mean ± SD, n = 2). (B) SPR analysis of K1K1 to immobilized MET receptor extracellular domain. The main plot shows the binding isotherm with equilibrium response plotted at different concentrations. The inserted graph shows the binding curves at different concentrations of K1K1. (C) SPR analysis of NK1 to immobilized MET receptor extracellular domain. As in (B), the main plot shows the binding isotherm and the inserted graph shows the binding curves at different concentrations of NK1. Source data are available for this figure.

Article Snippet: For dose–response analysis, 8-wk-old FVB mice weighing 19–21 g (Charles River) were i.p. injected with 0.1, 0.5, 1, and 5 μg of K1K1 diluted in 100 μl of PBS and euthanized per cervical dislocation 10 min after injection.

Techniques: Amplified Luminescent Proximity Homogenous Assay, Titration, Binding Assay

Journal: Life Science Alliance

Article Title: Dimerization of kringle 1 domain from hepatocyte growth factor/scatter factor provides a potent MET receptor agonist

doi: 10.26508/lsa.202201424

Figure Lengend Snippet: (A) MDCK cells were treated with 0.7 mM anisomycin for 16 h with the addition of hepatocyte growth factor/scatter factor, NK1, or different batches of K1K1 or K1K1H6 at different concentrations. Plotted are two different batches of K1K1 and three different batches of K1K1H6. Measurements are expressed in raw fluorescence signal as technical duplicates (mean ± SD, n = 2). (B) MDCK cell scattering at different concentrations of K1K1, K1K1S2, and K1K1S3 showing the lowest concentration at which each protein is still active and the subsequent dilution at which no more scattering is observed. Hepatocyte growth factor/scatter factor and K1K1 1 nM were used as positive control (maximum scattering) and PBS as the negative control. For complete half-log dilution of agonist series, see source data. The scale bar represents 100 μm. Source data are available for this figure.

Article Snippet: For dose–response analysis, 8-wk-old FVB mice weighing 19–21 g (Charles River) were i.p. injected with 0.1, 0.5, 1, and 5 μg of K1K1 diluted in 100 μl of PBS and euthanized per cervical dislocation 10 min after injection.

Techniques: Fluorescence, Concentration Assay, Positive Control, Negative Control

Journal: Life Science Alliance

Article Title: Dimerization of kringle 1 domain from hepatocyte growth factor/scatter factor provides a potent MET receptor agonist

doi: 10.26508/lsa.202201424

Figure Lengend Snippet: (A) MDCK cell scattering at different concentrations K1K1, K1K1S2, and K1K1S4 showing the lowest concentration at which each protein is still active and the subsequent dilution at which no more scattering is observed. Hepatocyte growth factor/scatter factor (HGF/SF) is used as positive control to generate maximum scattering. For complete half-log dilution of agonist series, see Data Source Image File. (B) 3D reconstruction by z-stacking of fluorescence microscopy images taken of large MDCK cell colonies stimulated with 100 pM HGF/SF or 10 nM K1K1 and mutants for 4 wk. The combined fluorescence of DAPI (red) and Evans blue staining (grey scale) shows the extensive branching morphogenesis and tubulogenesis induced by both proteins. (C) Boyden chamber migration assay using SKOV3 cells. Three independent assays were performed in which SKOV3 cells were treated for 6 h with indicated concentrations of HGF/SF, K1K1, K1K1S2, and K1K1S4. 0.1 nM HGF/SF was taken as 100% to which all other conditions where compared. Statistical significance was calculated with ANOVA followed by Dunnett’s test with P < 0.05 and * indicates significantly difference. Error bars represent mean ± SD based (n = 5). Source data are available for this figure.

Article Snippet: For dose–response analysis, 8-wk-old FVB mice weighing 19–21 g (Charles River) were i.p. injected with 0.1, 0.5, 1, and 5 μg of K1K1 diluted in 100 μl of PBS and euthanized per cervical dislocation 10 min after injection.

Techniques: Concentration Assay, Positive Control, Fluorescence, Microscopy, Staining, Migration

Journal: Life Science Alliance

Article Title: Dimerization of kringle 1 domain from hepatocyte growth factor/scatter factor provides a potent MET receptor agonist

doi: 10.26508/lsa.202201424

Figure Lengend Snippet: (A) MDCK cell (∼500) were seeded into a thick layer of a 1:1 collagen/Growth Factor Reduced Matrigel and treated with semi-log dilutions (100 pM to 300 nM) of hepatocyte growth factor/scatter factor, K1K1, K1K1S2, and K1K1S4 twice a week for 1 mo. The cells were fixed and stained with Evans blue and DAPI. The image of the full plate is presented. (B) Colonies were observed in bright field using inverted microscope observed 4× objective on a Nikon Eclipse TS100 microscope. (C) Phosphorylation analysis of MET signalling pathway by Western blot on HeLa cell lysates after stimulation with semi-log dilution (0.3–30 nM) of K1K1, K1K1S2, or K1KS4 for 10 min. Loading controls are based on total MET, total Akt, and total ERK present in each lane. Source data are available for this figure.

Article Snippet: For dose–response analysis, 8-wk-old FVB mice weighing 19–21 g (Charles River) were i.p. injected with 0.1, 0.5, 1, and 5 μg of K1K1 diluted in 100 μl of PBS and euthanized per cervical dislocation 10 min after injection.

Techniques: Staining, Inverted Microscopy, Microscopy, Phospho-proteomics, Western Blot

Journal: Life Science Alliance

Article Title: Dimerization of kringle 1 domain from hepatocyte growth factor/scatter factor provides a potent MET receptor agonist

doi: 10.26508/lsa.202201424

Figure Lengend Snippet: 8-wk-old FVB mice were injected with PBS (Ctrl) or 5 μg of K1K1 either per I.V. or I.P. route of injection. Mice were euthanized 10 min after injection after which MET, Akt, and ERK phosphorylation in liver homogenate was determined by Western blot. Blot present total ERK and Akt proteins as loading controls. Source data are available for this figure.

Article Snippet: For dose–response analysis, 8-wk-old FVB mice weighing 19–21 g (Charles River) were i.p. injected with 0.1, 0.5, 1, and 5 μg of K1K1 diluted in 100 μl of PBS and euthanized per cervical dislocation 10 min after injection.

Techniques: Injection, Phospho-proteomics, Western Blot

Journal: Life Science Alliance

Article Title: Dimerization of kringle 1 domain from hepatocyte growth factor/scatter factor provides a potent MET receptor agonist

doi: 10.26508/lsa.202201424

Figure Lengend Snippet: (A) 8-wk-old FVB mice were injected with PBS (Ctrl) or different amounts of K1K1 after which MET, AKT, and ERK phosphorylation in liver homogenate was determined using Western blot. Mice were euthanized 10 min after injection. Results are presented as experimental duplicate (n = 2). (B) MET, AKT, and ERK phosphorylation were detected by Western blot at different time points after injection of 5 μg of K1K1. Results are presented as experimental duplicate (n = 2) except for control and 10 min conditions. Both blots present total MET, Akt, and ERK proteins as loading controls. Source data are available for this figure.

Article Snippet: For dose–response analysis, 8-wk-old FVB mice weighing 19–21 g (Charles River) were i.p. injected with 0.1, 0.5, 1, and 5 μg of K1K1 diluted in 100 μl of PBS and euthanized per cervical dislocation 10 min after injection.

Techniques: Injection, Phospho-proteomics, Western Blot, Control

Journal: Life Science Alliance

Article Title: Dimerization of kringle 1 domain from hepatocyte growth factor/scatter factor provides a potent MET receptor agonist

doi: 10.26508/lsa.202201424

Figure Lengend Snippet: (A) Hematoxylin-erythrosin B staining of mouse livers submitted to an adapted Lieber DeCarli model (20× magnification). Analyses were performed on 10 animals in control groups (Ctr + vehicle and Ctr +10 μg K1K1) and 15 animals in ethanol-treated groups (LDC ± K1K1). Scale bar = 50 μm. (B) Steatosis was quantified in each mouse liver using a steatosis score (see STAR Methods). (C) Results are expressed as mean ± SD (C). The mRNA expression of triglyceride metabolism markers (ApoB, PPARa, and LDLR) was analysed in mouse livers using RT-qPCR with b-actin as housekeeping gene. Results are expressed in relative units (RU) and represented as mean ± SD. Source data are available for this figure.

Article Snippet: For dose–response analysis, 8-wk-old FVB mice weighing 19–21 g (Charles River) were i.p. injected with 0.1, 0.5, 1, and 5 μg of K1K1 diluted in 100 μl of PBS and euthanized per cervical dislocation 10 min after injection.

Techniques: Staining, Control, Expressing, Quantitative RT-PCR

Journal: Life Science Alliance

Article Title: Dimerization of kringle 1 domain from hepatocyte growth factor/scatter factor provides a potent MET receptor agonist

doi: 10.26508/lsa.202201424

Figure Lengend Snippet: (A) Hematoxylin-erythrosin B staining of mouse livers submitted to an adapted Lieber DeCarli model (×10). Analyses were performed on 10 animals in control groups (Ctrl + vehicle and Ctrl +10 μg K1K1) and 15 animals in ethanol-treated groups (LDC ± K1K1). Scale bar = 100 μm. (B) The mRNA expression of MET and inflammatory cytokines TNFα and IL-6 was analyzed in mouse livers using RT-qPCR with b-actin as the housekeeping gene. Results are expressed relative unit (RU) and represented as mean ± SD. Source data are available for this figure.

Article Snippet: For dose–response analysis, 8-wk-old FVB mice weighing 19–21 g (Charles River) were i.p. injected with 0.1, 0.5, 1, and 5 μg of K1K1 diluted in 100 μl of PBS and euthanized per cervical dislocation 10 min after injection.

Techniques: Staining, Control, Expressing, Quantitative RT-PCR

Journal: Life Science Alliance

Article Title: Dimerization of kringle 1 domain from hepatocyte growth factor/scatter factor provides a potent MET receptor agonist

doi: 10.26508/lsa.202201424

Figure Lengend Snippet: (A) NK1 and K1K1 were each injected at 2 and 1 μM on a heparin-coated SC HEP0320.a chip (Xantec). (B) Comparison of heparin affinity of K1K1, K1K1S2, and K1K1S4, injected at 2.5 and 1.25 μM each. All injections were done with a flow of 30 μl/min at 25°C in PBS running buffer.

Article Snippet: For dose–response analysis, 8-wk-old FVB mice weighing 19–21 g (Charles River) were i.p. injected with 0.1, 0.5, 1, and 5 μg of K1K1 diluted in 100 μl of PBS and euthanized per cervical dislocation 10 min after injection.

Techniques: Injection, Comparison