Journal: Biology

Article Title: Chemical Composition, Antioxidant Capacity, and Anticancerous Effects against Human Lung Cancer Cells of a Terpenoid-Rich Fraction of Inula viscosa

doi: 10.3390/biology13090687

Figure Lengend Snippet: IVL DCM decreased the viability of A549 cells. ( A ) MTT assay was used to measure the viability of A549 and HDFn cells treated with the indicated concentrations of IVL DCM for 24, 48, or 72 h. Data represents the mean ± SEM of the percent viability of treated cells compared to vehicle-treated cells. Data are the mean of three independent experiments ( n = 3). *** p < 0.001, **** p < 0.0001. ( B ) A549 cells treated with the indicated concentrations of IVL DCM for 24 h were stained with DAPI nuclear stain. DAPI fluorescence was imaged at 20X magnification using a BioTek Cytation 5 reader. ( C ) A549 cells were treated with the indicated concentrations of IVL DCM for 24 h and then stained with crystal violet. The micrographs represent stained cells imaged at 20X magnification by light microscopy using the Invitrogen EVOS ® FL Cell Imaging System.

Article Snippet: A549 human lung adenocarcinoma cells, the most common subtype of NSCLC, which accounts for approximately 85% of lung cancer cases [ ], human normal neonatal fibroblasts (HDFn), SK-OV-3 (human ovarian cancer), MCF-7 (human breast cancer), MDA-MB-231 (human breast cancer), HepG2 (human liver cancer), and HCT116 (human colorectal cancer) cells were obtained from ATCC and cultured at 37 °C and 5% CO 2 in a humidified incubator.

Techniques: MTT Assay, Staining, Fluorescence, Light Microscopy, Imaging

Journal: Frontiers in Cellular Neuroscience

Article Title: Reprogramming of adult human dermal fibroblasts to induced dorsal forebrain precursor cells maintains aging signatures

doi: 10.3389/fncel.2023.1003188

Figure Lengend Snippet: Phase contrast images of (A) HDFs and (B) resulting hiDFP colonies from 20/30-, 50- and 70-year-old subjects. Scale = 300 μm. HDF, human dermal fibroblast; hiDFP, human induced dorsal forebrain precursor.

Article Snippet: Nine adult human dermal fibroblast cell lines (HDFs) were obtained from Cell Applications Inc or The NINDS Human Cell and Data Repository.

Techniques:

Journal: Frontiers in Cellular Neuroscience

Article Title: Reprogramming of adult human dermal fibroblasts to induced dorsal forebrain precursor cells maintains aging signatures

doi: 10.3389/fncel.2023.1003188

Figure Lengend Snippet: Reprogrammed hiDFPs maintain aging-associated features. (A) SA-βgal activity assay of HDFs and hiDFPs. No statistical significance was determined by two-way mixed ANOVA. (B) Mitochondrial superoxide levels in HDFs and hiDFPs. Data represent mean ± SEM with n = 3. Simple main effect of reprogramming on mitochondrial superoxide levels determined by two-way mixed ANOVA with * for p < 0.05.

Article Snippet: Nine adult human dermal fibroblast cell lines (HDFs) were obtained from Cell Applications Inc or The NINDS Human Cell and Data Repository.

Techniques: Activity Assay

Journal: Frontiers in Cellular Neuroscience

Article Title: Reprogramming of adult human dermal fibroblasts to induced dorsal forebrain precursor cells maintains aging signatures

doi: 10.3389/fncel.2023.1003188

Figure Lengend Snippet: Reprogrammed hiDFPs exhibit increased amounts of DNA methylation as measured by the production of 5-methylcytosine (5-mC). (A) Amount of methylated DNA in hiPSCs and hiDFPs relative to HDF. Significance was determined by one-way ANOVA with Tukey post-hoc test. (B) Amount of methylated DNA in HDFs and hiDFPs. A simple main effect of reprogramming irrespective of age group was determined by two-way mixed ANOVA. Data represent mean ± SEM with n = 3. * for p < 0.05, ** for p ≤ 0.01.

Article Snippet: Nine adult human dermal fibroblast cell lines (HDFs) were obtained from Cell Applications Inc or The NINDS Human Cell and Data Repository.

Techniques: DNA Methylation Assay, Methylation

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: The Induction of Apoptosis in A375 Malignant Melanoma Cells by Sutherlandia frutescens

doi: 10.1155/2016/4921067

Figure Lengend Snippet: The effect of S. frutescens extract on the viability of (a) A375 melanoma, (b) Colo-800 melanoma, (c) HDF α fibroblast, and (d) Hek 293 cells after 24, 48, and 72 h of treatment. The viability was assessed using the alamarBlue assay and expressed as percentage relative to the control-treated cells (0 mg/mL). Error bars represent the SEM ( n ≥ 3) and ∗ indicates a significant difference from 0 mg/mL ( P < 0.05).

Article Snippet: The human primary dermal fibroblast cell line (HDF α ) was purchased from ScienCell, CA, USA.

Techniques: Alamar Blue Assay, Control

Journal: bioRxiv

Article Title: Protein kinase D1 phosphorylates CBX8 to facilitate the disassociation of PRC1 complex from p16 promoter and promotes cell senescence

doi: 10.1101/2020.08.09.242891

Figure Lengend Snippet: (A) ER: RAS IMR90 cells were induced senescent by 4-OHT treatment for 6 days, and then western blot was performed using the indicated antibodies. (B) PHBLV vector, FLAG-WT-CBX8 or FLAG-CBX8-S2A mutant were expressed in RAS-IMR90 cells with a lentiviral expression system. Then cells were subjected to western blot for the indicated proteins. (C) Total RNAs were extracted from (B) cells, then p16 mRNA level was measured by qPCR. (D) RAS-IMR90 cells infected with the indicated vectors were subjected to SA-β-gal activity staining. The percentages of SA-β-gal staining positive cells were statistically analyzed. Data are presented as mean ± s.d. from three independent experiments with 300 cells per experiment. (E) Growth curves of RAS-IMR90 cells infected with the indicated vectors were determined by CCK-8 assay. Data are presented as mean ± s.d. from three independent experiments, each performed in triplicate. (F) RAS-IMR90 cells infected with the indicated vectors were subjected to EdU incorporation assay. The percentages of EdU incorporating positive cells were statistically analyzed. Data are presented as mean ± s.d. from three independent experiments with 300 cells per experiment. (G) RAS-IMR90 cells infected with the indicated vectors were cultured for 14 days, then colony formation assay was performed by crystal violet staining cells. The representative magnified images (x40) were shown. (H-M) RAS-IMR90 cells were infected with HA-PKD1.CA lentiviral vector, then transfected with or without CBX8-S2A. Then cells were subjected to western blot (H), Realtime qPCR (I), SA-β-gal activity staining (J), cell growth assay (K), EdU incorporation assay (L), and colony formation assay (M), to evaluate cell senescent status.

Article Snippet: IMR90 human diploid fibroblasts (HDFs) were purchased from the American Type Culture Collection (ATCC) and cultured in DMEM with 10% FBS.

Techniques: Western Blot, Plasmid Preparation, Mutagenesis, Expressing, Infection, Activity Assay, Staining, CCK-8 Assay, Cell Culture, Colony Assay, Transfection, Growth Assay

Journal: bioRxiv

Article Title: Protein kinase D1 phosphorylates CBX8 to facilitate the disassociation of PRC1 complex from p16 promoter and promotes cell senescence

doi: 10.1101/2020.08.09.242891

Figure Lengend Snippet: (A) HEK293T cells were transfected with FLAG-WT-CBX8, CBX8-S2A or CBX8-S2E. Then IP assay was carried out using anti-FLAG antibody followed by immunoblotting with the indicated antibodies. (B) HEK293T cells were co-transfected with FLAG-WT-CBX8 or FLAG-CBX8-S2A mutant, with or without HA-PKD1.CA. Then IP assay was carried out using anti-FLAG antibody followed by immunoblotting with the indicated antibodies. (C) IMR90 cells co-transfected FLAG-CBX8 with or without HA-PKD1.CA, then IP assay was carried out using anti-FLAG antibody followed by immunoblotting with the indicated antibodies. (D) RAS-IMR90 cells were transfected with FLAG-WT-CBX8 or FLAG-CBX8-S2A mutant and induced to senescence, then IP assay was carried out using anti-FLAG antibody followed by immunoblotting with the indicated antibodies. (E) HEK 293T cells were transfected with CBX8-siRNA#1 or si-UTR, and CBX8 protein level was detected by western blot. (F) HepG2 cells were transfected with CBX8 UTR siRNA, then infected with FLAG-WT-CBX8, CBX8-S2A mutant or CBX8-S2E mutant. Then cells were subjected to western blot for the indicated proteins. (G) HepG2 cells were transfected with CBX8 UTR siRNA, then infected with FLAG-WT-CBX8 or FLAG-CBX8-S2A mutant, and with or without HA-PKD1.CA. Then cells were subjected to western blot for the indicated proteins. (H) HepG2 cells were transfected with FLAG-WT-CBX8, CBX8-S2A or CBX8-S2E. Then ChIP assay was performed using anti-FLAG antibody to measure the binding activity of WT-CBX8 and its mutants to the INK4A/ARF locus promoter.

Article Snippet: IMR90 human diploid fibroblasts (HDFs) were purchased from the American Type Culture Collection (ATCC) and cultured in DMEM with 10% FBS.

Techniques: Transfection, Western Blot, Mutagenesis, Infection, Binding Assay, Activity Assay

Journal: Molecules

Article Title: Volatile Natural Deep Eutectic Solvents (VNADESs) for Extraction of Shikonin Derivatives from Echium vulgare Roots and Evaluation of Biological Activity

doi: 10.3390/molecules31091434

Figure Lengend Snippet: Assessment of resazurin reduction in fibroblasts (HDFs) for ( a ) hexane (bars in yellow) and ( b ) TBa (bars in red) extracts. Control cells (green bars) without the addition of test samples, for which viability was assumed to be 100%. Data are presented as mean ± SD ( n = 3). Statistical analysis was performed using one-way ANOVA followed by Dunnett’s multiple comparisons test. Differences were considered statistically significant at p < 0.05 (*), p < 0.01(**), p < 0.001(***), relative to the control.

Article Snippet: Human dermal fibroblasts (HDFs) were obtained from CLS Cell Lines Service (Eppelheim, Germany) and were maintained in Dulbecco’s Modified Eagle’s Medium (DMEM; Biological Industries, Cromwell, CO, USA) supplemented with sodium pyruvate, L-glutamine, glucose (4.5 g/L), and 10% fetal bovine serum (FBS; Genos, Łódź, Poland).

Techniques: Control