Journal: International Journal of Molecular Sciences

Article Title: Novel Osteoblastogenic Activity of Magnolia kobus : The Pharmacological Potential for Osteoporosis

doi: 10.3390/ijms27052472

Figure Lengend Snippet: Effect of ME and magnolin on osteoblast differentiation in MC3T3-E1 cells. ( A ) Cell viability, ( B , C ) cell proliferation, ( D ) ALP staining, and ( E ) ALP activity assays were performed as described in Materials and Methods. Cells were treated with ME (left panel, 25–400 μg/mL) or magnolin (right panel, 1–40 μM) for 24 h ( A , B ) and 21 days ( C ), respectively. Untreated cells were cultured in serum-free MEM alpha ( A , B ) or 10% FBS-MEM alpha ( C – E ). For the differentiation experiments, cells were cultured in differentiation medium (DM: 10% FBS-MEM alpha containing 10 mM β-glycerophosphate and 50 µg/mL ascorbic acid) and treated with ME (25–100 μg/mL) or magnolin (1–10 μM) for 14 days ( D , E ). ( D ) ALP staining images were photographed using a microscope (original magnification ×40). Results are presented as the percentage ( A – C ) or the fold-increase ( E ) of untreated controls. Statistical significance is indicated ( # p < 0.05, ## p < 0.01, ### p < 0.001, compared with untreated cells; * p < 0.05, ** p < 0.01, *** p < 0.001, compared with 10% FBS- or DM-treated cells).

Article Snippet: MC3T3-E1 subclone 4 (CRL-2593) and RAW264.7 (TIB-71) cells were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Staining, Activity Assay, Cell Culture, Microscopy

Journal: International Journal of Molecular Sciences

Article Title: Novel Osteoblastogenic Activity of Magnolia kobus : The Pharmacological Potential for Osteoporosis

doi: 10.3390/ijms27052472

Figure Lengend Snippet: Effect of ME and magnolin on mineralization in MC3T3-E1 cells. ( A ) ARS staining and ( B ) mineralization assays were performed as described in Materials and Methods. Cells were cultured in DM and treated with ME (upper panel, 25–100 μg/mL) or magnolin (lower panel, 1–10 μM) for 21 days. Cell culture medium for untreated controls was 10% FBS-MEM alpha. ( A ) ARS staining images were photographed using a microscope (original magnification ×40). Results are presented as the fold-increase of untreated controls. Statistical significance is indicated ( ### p < 0.001, compared with untreated cells; ** p < 0.01, *** p < 0.001, compared with DM-treated cells).

Article Snippet: MC3T3-E1 subclone 4 (CRL-2593) and RAW264.7 (TIB-71) cells were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Staining, Cell Culture, Microscopy

Journal: International Journal of Molecular Sciences

Article Title: Novel Osteoblastogenic Activity of Magnolia kobus : The Pharmacological Potential for Osteoporosis

doi: 10.3390/ijms27052472

Figure Lengend Snippet: Effect of ME and magnolin on osteoblastogenesis-related transcription factors in MC3T3-E1 cells. Western blot analysis was performed as described in Materials and Methods. Cells were cultured in DM and treated with ( A ) ME (25–100 μg/mL) or ( B ) magnolin (1–10 μM) for 24 h. Cell culture medium for untreated controls was 10% FBS-MEM alpha. Results are presented as the fold-increase of untreated controls. Statistical significance is indicated ( ## p < 0.01, compared with untreated cells; * p < 0.05, ** p < 0.01, *** p < 0.001, compared with DM-treated cells).

Article Snippet: MC3T3-E1 subclone 4 (CRL-2593) and RAW264.7 (TIB-71) cells were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Western Blot, Cell Culture

Journal: International Journal of Molecular Sciences

Article Title: Novel Osteoblastogenic Activity of Magnolia kobus : The Pharmacological Potential for Osteoporosis

doi: 10.3390/ijms27052472

Figure Lengend Snippet: Effect of ME and magnolin on osteoblastogenic markers in MC3T3-E1 cells. Quantitative PCR analysis was performed as described in Materials and Methods. Cells were cultured in DM and treated with ( A ) ME (25–100 μg/mL) or ( B ) magnolin (1–10 μM) for 7 days. Untreated cells were cultured in 10% FBS-MEM alpha. GAPDH was used as control gene. Results are shown as the fold-increase of untreated controls. Statistical significance is indicated ( # p < 0.05, ## p < 0.01, ### p < 0.001, compared with untreated cells; * p < 0.05, ** p < 0.01, *** p < 0.001, compared with DM-treated cells).

Article Snippet: MC3T3-E1 subclone 4 (CRL-2593) and RAW264.7 (TIB-71) cells were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Real-time Polymerase Chain Reaction, Cell Culture, Control

Journal: International Journal of Molecular Sciences

Article Title: Novel Osteoblastogenic Activity of Magnolia kobus : The Pharmacological Potential for Osteoporosis

doi: 10.3390/ijms27052472

Figure Lengend Snippet: Effect of ME and magnolin on MAPK signaling pathways in MC3T3-E1 cells. Cells were cultured in DM and treated with ( A ) ME (25–100 μg/mL) or ( B ) magnolin (1–10 μM) for 15 min. Untreated cells were cultured in 10% FBS-MEM alpha. Results are shown as the fold-increase of untreated controls. Statistical significance is indicated ( # p < 0.05, ## p < 0.01, ### p < 0.001, compared with untreated cells; * p < 0.05, ** p < 0.01, *** p < 0.001, compared with DM-treated cells).

Article Snippet: MC3T3-E1 subclone 4 (CRL-2593) and RAW264.7 (TIB-71) cells were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Protein-Protein interactions, Cell Culture

Journal: International Journal of Molecular Sciences

Article Title: Novel Osteoblastogenic Activity of Magnolia kobus : The Pharmacological Potential for Osteoporosis

doi: 10.3390/ijms27052472

Figure Lengend Snippet: Involvement of p38 MAPK in ME- and magnolin-induced mineralization in MC3T3-E1 cells. ( A ) ARS staining and ( B ) mineralization assays were performed as described in Materials and Methods. Cells were cultured in DM and treated with ME (100 μg/mL) or magnolin (10 μM) for 21 days in the presence or absence of PD98059 (25 μM) and SB203580 (5 μM). Untreated cells were cultured in 10% FBS-MEM alpha. ( A ) ARS staining images were photographed using a microscope (original magnification ×40). Results are presented as the fold-increase of untreated controls. Statistical significance is indicated ( ### p < 0.001, compared with untreated cells; *** p < 0.001, compared with DM-treated cells).

Article Snippet: MC3T3-E1 subclone 4 (CRL-2593) and RAW264.7 (TIB-71) cells were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Staining, Cell Culture, Microscopy

Journal: Applied and Environmental Microbiology

Article Title: An RTCA-based assay as an innovative approach for thermal inactivation studies of hepatitis A virus

doi: 10.1128/aem.01822-25

Figure Lengend Snippet: Quantification of infectious HAV using RTCA-based assay. ( A ) Kinetic curves of CI values in FRhK-4 cells uninfected (blue, “mock”: cells treated with virus-free infection medium) or infected with different concentrations of HAV (from 10 5 PFU/100 µL [black] to 0.1 PFU/100 µL [pink]). The red arrow indicates the time at which HAV was added to sub-confluent monolayers of FRhK-4 cells. ( B ) Calibration curve between tCI 50 and log 10 of HAV concentration. Values of tCI 50 are presented as the time (hours post-seeding) at which a 50% decrease in CI values is observed. The straight line corresponds to the linear regression, and the dotted lines indicate the prediction interval at 95% and the confidence interval of the regression line.

Article Snippet: The FRhK-4 (fetal rhesus monkey kidney) cell line was purchased from the American Type Culture Collection (ATCC CRL-1688) (LGC standards SARL, Illkirch, France).

Techniques: Virus, Infection, Concentration Assay

Journal: Applied and Environmental Microbiology

Article Title: An RTCA-based assay as an innovative approach for thermal inactivation studies of hepatitis A virus

doi: 10.1128/aem.01822-25

Figure Lengend Snippet: Real-time impedance analysis of mock-infected (blue) and HAV-infected (red: inoculum; purple: heat-treated HAV) FRhK-4 cells at different incubation temperatures (37°C, 50°C, 56°C, 65°C, and 72°C and a dynamic range of 5°C–80°C). The incubation time is represented by a color gradient, with the longest times represented by the lightest colors. Representative data from one of three independent experiments are shown.

Article Snippet: The FRhK-4 (fetal rhesus monkey kidney) cell line was purchased from the American Type Culture Collection (ATCC CRL-1688) (LGC standards SARL, Illkirch, France).

Techniques: Infection, Incubation

Journal: bioRxiv

Article Title: A mRNA-LNP vaccine against Dengue Virus elicits robust, serotype-specific immunity

doi: 10.1101/2021.01.05.425517

Figure Lengend Snippet: A) Schematic of the DENV genome and engineered mRNA construct. An mRNA encoding for the prM and ENV viral proteins was engineered with N-terminal signal peptide sequence, 5’ and 3’ untranslated regions (UTR) flanking the coding sequence, a 3’ poly-A tail, and a 5’ cap-1 structure. In vitro synthesized mRNA is encapsulated in a lipid nanoparticle for use in in vitro and in vivo experiments. B) 293T cells were transfected with the in vitro transcribed mRNA encoding for the wild-type sequence (WT), or a mutant version with amino acid substitutions in the fusion-loop epitope (ΔFL). Lysate was analyzed by western blot with the domain III specific 1A1D-2 monoclonal antibody and the fusion-loop specific 4G2 monoclonal antibody. C) Supernatant from transfected cells was purified and concentrated through ultracentrifugation and analyzed for VLPs by western blots with the 1A1D-2 monoclonal antibody or anti-GAPDH. Unpurified cell lysate from WT mRNA transfected cells is included as a control. Shown are representative blots. D) Electron microscopy image of VLPs from purified supernatant of transfected 293T cells showing homogenous shape and size of approximately 30nm.

Article Snippet: Membranes were blotted with envelope domain III specific 1A1D-2 (1:600) monoclonal antibody (CDC Arbovirus Reference Collection) or envelope fusion-loop specific 4G2 (3.33 mg/ml) (BEI Cat# NR-50327, Novus Biologicals Cat# NBP2-52709FR).

Techniques: Construct, Sequencing, In Vitro, Synthesized, In Vivo, Transfection, Mutagenesis, Western Blot, Purification, Control, Electron Microscopy

Journal: Nature Communications

Article Title: In vivo PAR-CLIP (viP-CLIP) of liver TIAL1 unveils targets regulating cholesterol synthesis and secretion

doi: 10.1038/s41467-023-39135-8

Figure Lengend Snippet: a Schematic representation of in vivo PAR-CLIP (viP-CLIP) applied in mice. Mice are injected 6 times with 4SU over the time course of 12.5 h and sacrificed 15 h after the first dose. Intact organs are flash-frozen, grinded, and cross-linked with UV light (365 nm) under liquid nitrogen cooling. After cell lysis of the tissue powder, the RNA-RBP complexes are immunoprecipitated in presence of RNase, radiolabeled and the recovered RNA in the size of 19–35 nt is ligated to sequencing adapter, and used for cDNA library preparation and sequencing. b 4SU incorporation rates in tissues ( n = 3) after injection regime displayed in ( a ). HEK293 ( n = 3) and primary hepatocytes ( n = 3) were cultured in the presence of 100 mM 4SU for 16 h and subjected together with the tissue samples to HPLC analysis of 4SU incorporation rates. Values are mean +/– S.D. c Phosphorimage of urea-PAGE transferred to a nitrocellulose membrane that resolved 32 P-labeled RNA-TIAL1 complexes after immunoprecipitation of TIAL1 in several tissues. d Immunoblot showing tissue distribution of TIAL1 in mouse organs. e Autoradiograph of recovered RNAs as displayed in ( c ), after proteinase K treatment and 15% urea gel electrophoresis. 5’ radiolabeled synthetic RNAs of 19 and 35 nt length served as size markers. c , d , e Representative images of three independent replicates. f Graphic presentation of viP-CLIP cDNA library composition by RNA categories from recovered RNA from ( e ). Source data are provided as a Source Data file.

Article Snippet: HPLC purified ( ≥ 98%) 4-thiouridine (4SU) was purchased from Tocris (Cat No. 7005), protected against light, and dissolved fresh before use.

Techniques: In Vivo, Injection, Lysis, Immunoprecipitation, Sequencing, cDNA Library Assay, Cell Culture, Membrane, Labeling, Western Blot, Autoradiography, Nucleic Acid Electrophoresis