Journal: International Journal of Dentistry

Article Title: Acinar Cell Proliferation Promoted by BMP2 in Injured Mouse Parotid Gland: BMP2 Promotes Cell Proliferation in Parotid Gland

doi: 10.1155/2023/1765317

Figure Lengend Snippet: Mouse parotid acinar cells in primary culture. (a) Cell proliferative capacity of BMP2-added (100 ng/mL) and nonadded (control) groups after 48 hr of culture. The proliferative capacity of BMP2-added cultured cells was significantly increased compared to that in the control group. ∗∗ Indicates significance at P < 0.01, ∗ P < 0.05. Data were shown as mean ± SD. Three independent experiments were performed. (b) Protein expression of E-cadherin (an epithelial marker) and vimentin (a mesenchymal marker) in cultured cells of each group at 48 hr after the addition of BMP2. NIH3T3 (3T3) was used as a positive control for mesenchymal markers. RS: rat serum.

Article Snippet: NIH3T3 whole cell lysate (Novus Biologicals, Centennial, CO, USA) was used as a positive control for mesenchymal markers.

Techniques: Control, Cell Culture, Expressing, Marker, Positive Control

Journal: Cell calcium

Article Title: Ca 2+ -dependent binding of S100A6 to cofilin-1 regulates actin filament polymerization-depolymerization dynamics.

doi: 10.1016/j.ceca.2021.102457

Figure Lengend Snippet: Fig. 1. Interaction of S100A6 with cofilin-1 in NIH3T3 fibroblasts. (A) Pull-down assay with the use of protein lysate from NIH3T3 cells and S100A6 affinity resin (upper panel) or empty resin (lower panel). Lanes: 1-input, 2-unbound fraction, 3-last wash, 4-first wash with 0.5 M NaCl, 5-last wash with 0.5 M NaCl, 6-first wash with 1 M NaCl, 7- last wash with 1 M NaCl, 8- elution in buffer containing EGTA. Fractions were analyzed by SDS-PAGE (15% gel) fol lowed by immunoblotting developed with anti- cofilin-1 antibody. (B) Co-immunoprecipitation of S100A6 with cofilin-1 from NIH3T3 cell lysate. 30 μg of protein lysate was used directly for immunoblotting (input; lane 1 in both upper and lower panel) and 2.5 mg of protein lysate was incubated with (upper panel) or without (control, lower panel) anti-S100A6 monoclonal antibody and then with protein A/G agarose. In both panels, lane 2 shows unbound fraction, lane 3-last wash and lane 4-elution. Proteins were identified by immunoblotting using anti- cofilin-1 antibody. (C) Presence of S100A6- cofilin-1 complexes in NIH3T3 cells studied by PLA. Complexes of examined proteins are visualized in red; cell nuclei, stained with DAPI, are in blue. Scale bar is 20 μm.

Article Snippet: For co-immunoprecipitation assays 2.5 mg of protein lysate from NIH3T3 cells, obtained using the Plasma Membrane Protein Extraction Kit (Abcam) according to the manufacturer’s instruction was incubated with protein A/G-Agarose (Santa Cruz Biotechnology) for 1 h at 4◦C, as described by Jurewicz et al. [31].

Techniques: Pull Down Assay, SDS Page, Western Blot, Immunoprecipitation, Incubation, Control, Staining

Journal: Molecular Nutrition & Food Research

Article Title: Therapeutic Modulation of Inflammatory Signaling via Physalis Peruviana ‐Derived Exosomes: A Targeted Effect on IKK/NFκB/STAT1

doi: 10.1002/mnfr.70458

Figure Lengend Snippet: Effect of exosomes from P. peruviana on cell viability. (A) RAW 264,7 cells were exposed to different concentration of exosomes. (B) NIH3T3 cells were treated with various concentrations of exosomes. Data are expressed as mean ± SD. ( n = 4 biological replicates) * p < 0.05, ** p < 0.01, *** p < 0.001 compared to Ct (untreated control), using one‐way ANOVA followed by Tukey's multiple comparison test.

Article Snippet: NIH3T3 were acquired from ATCC (ATCC No. CRL‐1658.2) were subculture at a density of 3–5 × 10 3 cells/cm 2 in DMEM medium (Lonza) supplemented with 10% FBS, 1% penicillin‐streptomycin, and 4 mM L‐glutamine.

Techniques: Concentration Assay, Control, Comparison

Journal: Molecular Nutrition & Food Research

Article Title: Therapeutic Modulation of Inflammatory Signaling via Physalis Peruviana ‐Derived Exosomes: A Targeted Effect on IKK/NFκB/STAT1

doi: 10.1002/mnfr.70458

Figure Lengend Snippet: Antioxidant effect of P. peruviana exosomes in NIH3T3 cells. (A) Changes in cell viability after treatment with different concentrations of H 2 O 2 , (B) Cells treated with 500 µM H 2 O 2 and different concentrations of exosomes. Data are expressed as mean ± SD. ( n = 4 biological replicates) * p < 0.05, ** p < 0.01, *** p < 0.001 compared to Control (A) and to H 2 O 2 500 µM group (B), using one‐way ANOVA followed by Tukey's multiple comparison test.

Article Snippet: NIH3T3 were acquired from ATCC (ATCC No. CRL‐1658.2) were subculture at a density of 3–5 × 10 3 cells/cm 2 in DMEM medium (Lonza) supplemented with 10% FBS, 1% penicillin‐streptomycin, and 4 mM L‐glutamine.

Techniques: Control, Comparison

Journal: Molecular Nutrition & Food Research

Article Title: Therapeutic Modulation of Inflammatory Signaling via Physalis Peruviana ‐Derived Exosomes: A Targeted Effect on IKK/NFκB/STAT1

doi: 10.1002/mnfr.70458

Figure Lengend Snippet: Antioxidant enzymes expression levels following exposure to P. peruviana exosomes, H2O2 and exosomes + H2O2 in NIH3T3 cells. Data are expressed as mean ± SD. ( n = 3 biological replicates) * p < 0.05, ** p < 0.01, *** p < 0.001 compared to Control using one‐way ANOVA followed by Tukey's multiple comparison test. CAT, catalase; SOD1, superoxide dismutase 1; SOD2, superoxide dismutase 2; Cyt C ox, cytochrome c oxidase; GPx4, glutathione peroxidase 4.

Article Snippet: NIH3T3 were acquired from ATCC (ATCC No. CRL‐1658.2) were subculture at a density of 3–5 × 10 3 cells/cm 2 in DMEM medium (Lonza) supplemented with 10% FBS, 1% penicillin‐streptomycin, and 4 mM L‐glutamine.

Techniques: Expressing, Control, Comparison

Journal: Nucleic Acids Research

Article Title: Repeat-rich RNA guides repetitive genomic elements into biomolecular condensates for heterochromatin organization and muscle integrity

doi: 10.1093/nar/gkag168

Figure Lengend Snippet: CU-rich RNA promotes heterochromatin condensate organization during differentiation. ( A ) Nuclei of C2C12 myotubes (MT) treated with 1.5% 1,6-hexanediol (1,6-HD) or 1.5% 2,5-hexanediol (2,5-HD). Left: Representative images of 4,6-diamidino-2-phenylindole (DAPI)-stained nuclei. Scale bar, 5 μm. Middle: Time-course quantification of the number of heterochromatin foci per nucleus. Right: Boxplots show foci area (μm 2 , y -axis) at 5, 10, and 15 min posttreatment ( x -axis). n = 55 nuclei, three biological replicates. ( B ) Representative live-cell images of Hoechst 33342-stained MT nuclei before and after 1,6-HD treatment (0 and 15 min, respectively), taken from . Arrows indicate changes in heterochromatin foci intensity (pink, increased; blue, decreased), and the red arrow highlights an alteration in chromocenter integrity. ( C ) Number of heterochromatin foci per nucleus in MT following recovery from 1.5% 1,6-hexanediol (1,6-HD) treatment for 15 min, measured at indicated time points. n = 68 nuclei, three biological replicates. ( D ) Representative images of nuclei of myoblast (MB) and MT with or without 1,6-HD treatment (1.5%, 15 min). Right: Quantification of the number of heterochromatin foci per nucleus. n = 40 (MB), n = 60 (MT), three biological replicates. ( E ) Quantification of colocalization between indicated proteins and DAPI foci in MT with or without 1,6-HD treatment (1.5%, 15 min), by Pearson’s correlation coefficients. n = 60, three biological replicates. ( F ) Boxplot showing the distribution of Z -score of the interchromosomal interaction frequencies in MB and MT. ( G ) RNA FISH using ChRO1 and LacZ biotinylated probes. Biotin signal was detected by Fluorescein-conjugated Avidin DCS and amplified with biotinylated anti-Avidin and additional Fluorescein Avidin DCS. Right: Quantification of colocalization between biotin signal and DAPI foci. n = 50 nuclei. ( H ) Number of heterochromatin foci per nucleus of mouse fibroblast cells (NIH3T3) with or without doxycyline (Dox)-induced ChRO1a expression and/or 1,6-HD treatment (1.5%, 15 min). (EV; empty vector). n = 50, three biological replicates. ( I ) Number of heterochromatin foci per nucleus in MB with or without Dox-induced ChRO1a fragment (1–413, CUR) expression and/or 1,6-HD treatment (1.5%, 15 min). n = 75, three biological replicates. Statistical analyses and data presentation details are described in the “Materials and methods” section.

Article Snippet: C2C12 murine myoblast cells and NIH3T3 mouse fibroblast cells were obtained from the American-type culture collection and grown in a growth medium (GM) consisting of Dulbecco’s modified Eagle medium (DMEM) with 10% (v/v) fetal bovine serum at 37°C and 5% CO 2 .

Techniques: Staining, Avidin-Biotin Assay, Amplification, Expressing, Plasmid Preparation