Journal: Science Advances

Article Title: Decellularized zebrafish cardiac extracellular matrix induces mammalian heart regeneration

doi: 10.1126/sciadv.1600844

Figure Lengend Snippet: ( A to C ) Dual immunofluorescence detection and quantification of cTnT + /Ki67 + proliferating cardiomyocytes at 3 days after MI at the mid-infarct level of mouse left ventricles. Arrows indicate doubly positive cells. ( D to F ) Dual immunofluorescence detection and quantification of ErbB2 + /cTnT + cardiomyocytes at 3 days after MI at the mid-infarct level suggest the involvement of NRG1 signaling in zECM-treated groups. All image analyses were performed using 20 × 10–μm areas in five images of each heart ( n = 4 per group). All quantitative data represent means ± SD. *** P < 0.001 compared to mECM and saline; # P < 0.05 for hzECM versus nzECM. Scale bars, 50 μm.

Article Snippet: For ErbB2 inhibition in vivo, the ErbB2 inhibitor AG825 (sc-202045A, Santa Cruz Biotechnology) dissolved in DMSO was intraperitoneally injected once at 5 mg/kg immediately after the cardiac ECM administration ( , ).

Techniques: Immunofluorescence, Saline

Journal: Science Advances

Article Title: Decellularized zebrafish cardiac extracellular matrix induces mammalian heart regeneration

doi: 10.1126/sciadv.1600844

Figure Lengend Snippet: To inhibit ErbB2 activity in vivo, the ErbB2 inhibitor AG825 was intraperitoneally injected once (5 mg/kg) immediately after the administration of decellularized cardiac ECM. Cardiac contractile function is indicated by ( A ) fractional area change and ( B ) ejection fraction; LV dimension is indicated by ( C ) EDA and ( D ) ESA. No significant difference is observed between all groups at all time points ( n = 7 per group; all P > 0.05; data analyzed by two-way repeated-measures ANOVA). Dual immunofluorescence detection and quantification of ( E and F ) c-kit + /Ki67 + proliferating cardiac stem cells and ( G and H ) ErbB2 + /cTnT + cardiomyocytes. No significant difference is observed between all groups ( n = 4 per group, all P > 0.05). Scale bars, 50 μm.

Article Snippet: For ErbB2 inhibition in vivo, the ErbB2 inhibitor AG825 (sc-202045A, Santa Cruz Biotechnology) dissolved in DMSO was intraperitoneally injected once at 5 mg/kg immediately after the cardiac ECM administration ( , ).

Techniques: Activity Assay, In Vivo, Injection, Immunofluorescence

Journal: Cell structure and function

Article Title: miR-125b targets ARID3B in breast cancer cells.

doi: 10.1247/csf.11025

Figure Lengend Snippet: Fig. 4. (A) Western blot showing ERBB2 and pERK1/2 levels in SKBR3 (ERBB2 overexpressing positive control cell line), MCF7-EV and MCF7-125b cells. Blots were hybridized with anti-ERBB2, anti-pERK 1/2 and then with anti-β-actin antibodies to test equal loading. Image J program was used for the quantification of the bands. (B) AG825 effect on migration was determined by transwell migration assay. 200,000 pre-starved SKBR3 and MCF7 cells in 0.1 mL medium with 1% FBS and AG825 or DMSO (vehicle) were plated on the upper wells of migration chambers. Medium with 10% FBS was added to the lower wells. Cells were allowed to migrate for 24 h. 3 random fields were counted per membrane. 6 membrane replicates were used for each sample. * indicates statistical significance (p<0.05). “NS” indicates not significant.

Article Snippet: SKBR3 and MCF7 cells were pre-starved in McCoy’s 5A modified and MEM-Earle’s media with 1% FBS, respectively and after 24 hours, 200,000 cells in 0.1 mL medium containing 1% FBS and AG825 (50 μM, Tocris Bioscience) or DMSO were plated on the upper wells of transwell migration chambers.

Techniques: Western Blot, Positive Control, Migration, Transwell Migration Assay, Membrane

Journal: Pflugers Archiv

Article Title: Unlocking the potential: unveiling tyrphostins with Michael-reactive cyanoacrylate motif as promising inhibitors of human 5-lipoxygenase

doi: 10.1007/s00424-024-03019-7

Figure Lengend Snippet: Inhibition of 5-LO single cysteine mutants (C159S, C300S, C416S, or C418S) by degrasyn, tyrphostin A9, AG879, AG556, and CAPE normalized on DMSO control using 3 µg of recombinant 5-LO protein with the respective mutation. The inhibitor concentrations used correspond to the respective IC 50 value. Inhibitors were preincubated for 15 min in PBS/EDTA/ATP on ice before starting the reaction by the addition of 2 mM CaCl 2 and 10 µM AA for 10 min at 37 °C. Error bars depict standard deviation. Asterisks indicate significant changes towards r5-LO_WT with the same inhibitor treatment. *( P < 0.05), **( P < 0.01), ***( P < 0.001); ****( P < 0.0001), n = 3

Article Snippet: Tyrphostin A1, tyrphostin A9, AG18, AG30, ( E )-AG99, AG528, AG825, AG879, AG1024, degrasyn, RG13022, RG14620, SU1498, and WP1066 were obtained from MedChemExpress LLC (Monmouth Junction, NJ, USA).

Techniques: Inhibition, Control, Recombinant, Mutagenesis, Standard Deviation

Journal: Pflugers Archiv

Article Title: Unlocking the potential: unveiling tyrphostins with Michael-reactive cyanoacrylate motif as promising inhibitors of human 5-lipoxygenase

doi: 10.1007/s00424-024-03019-7

Figure Lengend Snippet: Mass spectra of r5-LO_WT or r5-LO_4C incubated with inhibitors (blue or red line). 20 µM r5-LO_WT (left) or r5-LO_4C (right) was incubated with 100 µM degrasyn ( A ), AG556 ( B ), tyrphostin A9 ( C ), AG879 ( D ), or CAPE ( E ) at RT for at least 1 h before analysis. Measurement was performed using an Agilent 1260 Infinity HPLC with an Agilent 6230 TOF LC/MS detector. Obtained data were processed using Mass Hunter BioConfirm (B.08.00), and resulted counts were normalized to the highest peak of the spectrum

Article Snippet: Tyrphostin A1, tyrphostin A9, AG18, AG30, ( E )-AG99, AG528, AG825, AG879, AG1024, degrasyn, RG13022, RG14620, SU1498, and WP1066 were obtained from MedChemExpress LLC (Monmouth Junction, NJ, USA).

Techniques: Incubation, Liquid Chromatography with Mass Spectroscopy

Journal: Molecular Cancer

Article Title: Yin Yang-1 suppresses invasion and metastasis of pancreatic ductal adenocarcinoma by downregulating MMP10 in a MUC4/ErbB2/p38/MEF2C-dependent mechanism

doi: 10.1186/1476-4598-13-130

Figure Lengend Snippet: Mechanisms involved in the tumor suppression role of YY1. (a) Effects of YY1 on MUC4, ErbB2, MEF2C, MMP10 and MAPK (Erk1/2, JNK and p38) signaling pathways were assayed by western blotting. BXPC-YY1 indicates YY1overexpressing BXPC-3 cells; BXPC-Vector indicates BXPC-3 cells transfected with a control vector; BXPC-YY1 shRNA indicates YY1 knockdown BXPC-3 cells; BXPC-Scramble shRNA indicates BXPC-3 cells transfected with a vector expressing Scramble shRNA. GAPDH was used as the internal control. (b) Bands were quantified by densitometry. Histograms of the ratio (phosphorylated/constitutive form) for Erk1/2, JNK and p38 MAPK kinases are shown. (c) BXPC-YY1 shRNA cells were transfected with MMP10 siRNA or negative control siRNA. 12 h after transfection, cell migration assays were performed. The upper chambers were seeded with various cell lines. The membranes of the chambers were stained with 0.1% crystal violet. Scale bar, 100 μm. (d) Quantification of the data from Figure c. OD values from three independent experiments were assessed. (e) Luciferase activities of MMP10 promoter-transfected BXPC-Scramble shRNA or BXPC-YY1 shRNA cells treated with MUC4 siRNA or various inhibitors of signal transduction molecules. MUC4 blockage, the ErbB2 inhibitor (AG825, 25 μM) and the p38 inhibitor (SB203580, 10 μM) significantly inhibited YY1 knockdown-stimulated luciferase activity in BXPC-YY1 shRNA cells transfected with the MMP10 promoter. (f) The YY1 knockdown-stimulated luciferase activity could be significantly inhibited by MEF2C blockage. Luciferase activity of MMP10 promoter in BXPC-YY1 shRNA cells was significantly decreased when the presumed MEF2C binding site (nucleotides −881 to −890) was mutated. (*represents p < 0.05, **represents p < 0.01, # represents p < 0.001, when compared to the control group).

Article Snippet: At 12 h after transfection, the medium was replaced and cells were treated for 24 h with 25 μM AG825 (Merck Millipore), 10 μM SB203580 (Cell Signaling Technology) or equivalent amounts of DMSO vehicle (Sigma) as a control.

Techniques: Protein-Protein interactions, Western Blot, Plasmid Preparation, Transfection, Control, shRNA, Knockdown, Expressing, Negative Control, Migration, Staining, Luciferase, Transduction, Activity Assay, Binding Assay