Journal: Cells

Article Title: Fixed-Bed Bioreactor Culture Enhances Yield and Reparative Properties of hTERT Mesenchymal Stem Cell Extracellular Vesicles

doi: 10.3390/cells15070654

Figure Lengend Snippet: EV function and signaling activity. ( a ) Primary keratinocytes were exposed to IR and treated with EVs from either flask or bioreactor cultures. Cell viability was assessed at 24, 48, and 72 h using the CellTiter-Glo assay. ( b ) Non-irradiated keratinocytes were treated with EVs from flask or bioreactor cultures, and viability was assessed after 72 h using the same assay. * = p < 0.05; *** = p < 0.001; **** = p < 0.0001. ( c ) Western blot of cell cycle proteins associated with hTERT MSC EVs produced from both the bioreactor and flask-based cultures. ( d ) In vitro kinase assay of EV-associated CDK2. EVs were immunoprecipitated with CDK2 antibodies or IgG control and kinase activity was assessed using Histone H1 as substrate.

Article Snippet: Normal primary epidermal keratinocytes (ATCC ® PCS-200-011TM) were maintained in Dermal Cell Basal Medium (ATCC ® PCS-200-030TM) supplemented with Keratinocyte Growth Kit (ATCC ® PCS-200-040TM).

Techniques: Activity Assay, Glo Assay, Irradiation, Western Blot, Produced, In Vitro, Kinase Assay, Immunoprecipitation, Control

Journal: Cell Death Discovery

Article Title: Trifluoperazine causes mast cell apoptosis through a secretory granule-mediated pathway

doi: 10.1038/s41420-026-03122-x

Figure Lengend Snippet: A Human lung smooth muscle cells (HLSMCs), lung fibroblasts (HLFs), and small airway epithelial cells (HSAECs) were treated with TFP at the indicated concentrations for 24 h. Human peripheral blood eosinophils and neutrophils were treated with TFP for 2 h. Cell viability was assessed by staining the cells with Annexin V (AnnV) and DRAQ7. Viable cells, AnnV − DRAQ7 − ; apoptotic cells, AnnV + DRAQ7 − ; necrotic/late apoptotic cells, AnnV + DRAQ7 + . HSAECs, n = 6 from three independent experiments; HLSMCs, eosinophils, neutrophils, n = 4 from four independent experiments; HLFs, n = 3 from one individual experiment representative of three independent experiments (One-way ANOVA for HLSMCs, HLFs; Kruskal-Wallis for HSAECs; Friedman test for eosinophils, neutrophils). B – C Bone marrow-derived MCs (BMMCs) and peritoneal cell-derived MCs (PCMCs) treated under the same conditions as in ( A ) for 24 h. BMMCs, n = 5 from two independent experiments; PCMCs, n = 3 from one individual experiment representative of three independent experiments (One-way ANOVA). Untreated (control) cells were used for statistical comparisons to all other groups in all figures. The bar charts show mean values + SEM or median + interquartile range. * P < 0.05; ** P < 0 .01; **** P < 0.0001. D Effect of TFP on DNA degradation. MCs were preincubated with bafilomycin A1 (Baf A1) (20 nM) for 2 h followed by treatment with TFP (10 μΜ) for 2 h. DNA was extracted from MCs and fragmentation was assessed by agarose gel electrophoresis. St standard marker.

Article Snippet: Primary human lung fibroblasts (HLFs) (PCS-201-013) and primary human lung smooth muscle cells (HLSMCs)(PCS-130-010) were obtained from American Type Culture Collection (ATCC, Manassas, VA) and cultured following the manufacturer’s instructions.

Techniques: Staining, Derivative Assay, Control, Agarose Gel Electrophoresis, Marker

Journal: International Journal of Molecular Sciences

Article Title: A Simple Ionic-Gelation Method for Chitosan Nanoparticle Synthesis and Standardized Protocols for Biological Safety Assessment: Antibacterial Activity, Phytotoxicity, and Biocompatibility

doi: 10.3390/ijms27083673

Figure Lengend Snippet: Cell viability of HDF cells treated with Ch NPs at concentrations of 2.5–160 µg/mL for 24 h. Viability was determined using the MTT assay and expressed as mean ± SE (n = 3). Different letters indicate statistically significant differences ( p < 0.05).

Article Snippet: The biocompatibility of Ch NPs was evaluated using primary HDF (ATCC ® PCS-201-012TM [ ], American Type Culture Collection, Manassas, VA, USA) and HaCaT cells (Cat. No. T0020001, AddexBio, San Diego, CA, USA).

Techniques: MTT Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: IFN-beta1a inhibits the secretion of Th17-polarizing cytokines in human dendritic cells via TLR7 up-regulation.

doi: 10.4049/jimmunol.0802226

Figure Lengend Snippet: FIGURE 2. IFN-1a treatment up-regulates TLR7 and its signaling pathway, while it down-regulates the expression of IL-1R and IL-1/. A, Three 106 DCs per condition derived from 10 RR MS patients were pretreated with IFN-1a for 24 h and then matured with LPS for 2 h before RNA extraction. TLR7, MyD88, IRAK4, TRAF6, IL-1R1, IL-1, and IL-1 gene expression was measured by RT- PCR. The results are presented as relative gene expression normalized for the 18S RNA expression. B, Three 106 DCs per condition from 16 RR MS patients were pretreated with IFN-1a for 24 h and then matured with LPS for 48 h. DCs were stained with PE-Cy-CD11c, allophycocyanin-CD1a mAb, and anti-IL-1R1 Ab, followed by FITC-conjugated secondary Ab, anti-TLR7 primary Ab, and FITC-conjugated secondary Ab. FITC-conjugated IgG isotype Ab was used as a control. The percentage of positive cells was determined in the CD1a-gated population. C, Three 106 DCs per condition derived from three RR MS patients were pretreated by IFN-1a for 24 h and then matured with LPS for 5 h before protein extraction. The expression of TLR7, MyD88, IRAK4, TRAF6, IL-1R1, and -actin were measured by Western blotting. The results present one of three similar experiments.

Article Snippet: Nonspecific binding was blocked for 1 h in the blocking buffer (10 mM Tris (pH 7.5), 150 mM NaCl, 2% fish gelatin, and 1% OVA in double-distilled water) and incubated overnight with rabbit anti-human Ab for TLR7 (Abcam), MyD88, TRAF6 (Santa Cruz Biotechnology), and IRAK4 (ProSci) and with mouse anti-human Ab for IL1R1 and -actin (Santa Cruz Biotechnology).

Techniques: Expressing, Derivative Assay, RNA Extraction, Gene Expression, Reverse Transcription Polymerase Chain Reaction, RNA Expression, Staining, Control, Protein Extraction, Western Blot

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: IFN-beta1a inhibits the secretion of Th17-polarizing cytokines in human dendritic cells via TLR7 up-regulation.

doi: 10.4049/jimmunol.0802226

Figure Lengend Snippet: FIGURE 4. IFN-1a modifies MyD88/IRAK4/ TRAF6 signaling and cytokine production in DCs through its induction of TLR7 expression. Three 106/well DCs generated from six RR MS patients were plated in antibiotic-free medium. After a 24-h incubation at 37oC, the DCs were transfected with TLR7 siRNA or control siRNA. The siRNA-treated cells were harvested and cultured in serum-free me- dium in the absence or presence of IFN-1 for 24 h, followed by LPS maturation for 5 h before the pro- tein extraction, and for 48 h before the SN collec- tion. A, The expression of TLR7, MyD88, IRAK4, TRAF6, IL-1R1, and -actin was measured by Western blotting. The results present one of three similar experiments. B, The production of IL-1, TGF-1, IL-23/p19, and IL-27 was measured by ELISA. The results present six experiments per- formed in triplicate. Statistical analysis was per- formed using a repeated measures ANOVA. , p 0.05; , p 0.01; and , p 0.001.

Article Snippet: Nonspecific binding was blocked for 1 h in the blocking buffer (10 mM Tris (pH 7.5), 150 mM NaCl, 2% fish gelatin, and 1% OVA in double-distilled water) and incubated overnight with rabbit anti-human Ab for TLR7 (Abcam), MyD88, TRAF6 (Santa Cruz Biotechnology), and IRAK4 (ProSci) and with mouse anti-human Ab for IL1R1 and -actin (Santa Cruz Biotechnology).

Techniques: Expressing, Generated, Incubation, Transfection, Control, Cell Culture, Extraction, Western Blot, Enzyme-linked Immunosorbent Assay