Journal: BMJ Open Ophthalmology

Article Title: Protective effects of blue light-blocking shades on phototoxicity in human ocular surface cells

doi: 10.1136/bmjophth-2018-000217

Figure Lengend Snippet: Representative fluorescent microscopic images of immunostained cells with anti-pan-keratin antibody and its isotype control antibody (mouse IgG1). (A, B) Cells from human corneal surface origin (peripheral portion) and normal human dermal fibroblasts immunostained with anti-pan-keratin antibody, respectively. (C, D) Cells from human corneal surface origin (peripheral portion) and normal human dermal fibroblasts immunostained with an isotype control antibody (mouse IgG1), respectively. Scale bar=100 µm.

Article Snippet: Normal human dermal fibroblasts (NHDF, DS Pharma Biomedical, Osaka, Japan) were used as a negative control.

Techniques: Control

Journal: bioRxiv

Article Title: Viral microRNA regulation of Akt is necessary for reactivation of Human Cytomegalovirus from latency in CD34 + hematopoietic progenitor cells and humanized mice

doi: 10.1101/2024.05.24.595672

Figure Lengend Snippet: (A, E) A dual luciferase reporter containing the 3’UTR of Akt (A) or Pin1 (E) was cotransfected into HEK293T cells along with miRNA mimics. Luciferase expression was assessed 24hrs post-transfection. The relative expression is shown as a percentage of Neg. Error bars represent the standard deviation from three separate experiments (**p<0.005 by unpaired t-test). (B) HEK293T cells were co-transfected with Akt-GFP and miRNA mimics or siRNA. Lysates were harvested 24hr post-transfection and immunoblotted for GFP and GAPDH. Quantification from one representative blot shows relative expression levels of Akt-GFP compared to Neg (normalized to GAPDH). (C) Quantification of (B) from three separate experiments (*p<0.05, **p<0.005, ***p<0.0005 [one-way ANOVA with Tukey’s multiple comparison test]). (D) NHDF cells were transfected with miRNA mimics, negative control (Neg), or siRNA. RNA has harvested 72hr post-transfection, and quantitative RT-PCR for Akt was performed. Expression levels were normalized to 18S and compared to Neg (*p<0.05, **p<0.005, ***p<0.0005 [one-way ANOVA with Tukey’s multiple comparison test]). (F) NHDF were transfected with miRNA mimics or siRNA. Lysates were harvested 72hr post-transfection and immunoblotted for Pin1, Akt, and GAPDH. Quantification from one representative blot shows relative expression levels of Akt compared to Neg (normalized to GAPDH). (G) Quantification of (F) from three separate experiments (*p<0.05, ***p<0.0005 [one-way ANOVA with Tukey’s multiple comparison test]).

Article Snippet: HEK293 and adult normal human dermal fibroblasts (NHDF) were obtained from ATCC and cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% heat-inactivated fetal bovine serum (FBS; Hyclone), 100 units/ml penicillin, 100 μg/ml streptomycin, and 100 μg/ml glutamine (Thermofisher).

Techniques: Luciferase, Expressing, Transfection, Standard Deviation, Comparison, Negative Control, Quantitative RT-PCR

Journal: bioRxiv

Article Title: Viral microRNA regulation of Akt is necessary for reactivation of Human Cytomegalovirus from latency in CD34 + hematopoietic progenitor cells and humanized mice

doi: 10.1101/2024.05.24.595672

Figure Lengend Snippet: (A,B) NHDF were infected with WT, ΔmiR-UL36/112/148D, or Mock infected for 48hr, serum starved overnight, and then stimulated +/−EGF for 15 minutes. Lysates were harvested and immunoblotted for Akt phosphorylated at T308 (A) or S473 (B) as well as total Akt, HCMV IE2, and GAPDH. Quantification from one representative blot shows relative expression levels of p-Akt compared to Neg (normalized to GAPDH). (C, D) Quantification of p-Akt levels from (A, B), respectively, from three separate experiments (comparing +EGF conditions *p<0.05, ***p<0.0005, ****p<0.0001 [two-way ANOVA with Tukey’s multiple comparison test]). (E, F) Ratio of p-Akt to total Akt levels in WT and ΔmiR-UL36/112/148D from (A, B), respectively, from three separate experiments, normalized to WT (*p<0.05 [unpaired t-test]).

Article Snippet: HEK293 and adult normal human dermal fibroblasts (NHDF) were obtained from ATCC and cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% heat-inactivated fetal bovine serum (FBS; Hyclone), 100 units/ml penicillin, 100 μg/ml streptomycin, and 100 μg/ml glutamine (Thermofisher).

Techniques: Infection, Expressing, Comparison

Journal: bioRxiv

Article Title: Viral microRNA regulation of Akt is necessary for reactivation of Human Cytomegalovirus from latency in CD34 + hematopoietic progenitor cells and humanized mice

doi: 10.1101/2024.05.24.595672

Figure Lengend Snippet: (A, B, E, F) NHDF were infected with WT, ΔmiR-UL36/112/148D, or Mock infected for 48hr, serum starved overnight, and then stimulated +/−EGF for 15 minutes. Lysates were harvested and immunoblotted for indicated phosphorylated and total proteins as well as HCMV IE2 and GAPDH. Quantification from one representative blot shows relative expression levels of p-protein compared to Neg (normalized to GAPDH). (C, D, G, H) Quantification of (A, B, E, F), respectively, from three separate experiments (comparing +EGF conditions, **p<0.005, ***p<0.0005, ****p<0.0001 [two-way ANOVA with Tukey’s multiple comparison test]).

Article Snippet: HEK293 and adult normal human dermal fibroblasts (NHDF) were obtained from ATCC and cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% heat-inactivated fetal bovine serum (FBS; Hyclone), 100 units/ml penicillin, 100 μg/ml streptomycin, and 100 μg/ml glutamine (Thermofisher).

Techniques: Infection, Expressing, Comparison

Journal: bioRxiv

Article Title: Viral microRNA regulation of Akt is necessary for reactivation of Human Cytomegalovirus from latency in CD34 + hematopoietic progenitor cells and humanized mice

doi: 10.1101/2024.05.24.595672

Figure Lengend Snippet: (A, B) NHDF were plated on coverslips and infected with WT, ΔmiR-UL36/112/148D, or Mock infected. Cells were fixed 72 hpi and stained for actin (phalloidin), FOXO3a, and nuclei (DAPI). (A) Representative images are shown. White dotted line shows outline of the nucleus in FOXO3a images. Scale bar, 20μM. (B) Image J software was used to quantify the average intensity of FOXO3a in the nucleus and the entire cell and graphed as a percentage of nuclear FOXO3a levels normalized to the whole cell. Error bars represent the standard error of the mean for 38-44 cells from each condition from three separate experiments (****p<0.0001 [one-way ANOVA with Tukey’s multiple comparison test]). (C-E) NHDF were infected with WT, ΔmiR-UL36/112/148D, or Mock infected for 72hr and RNA has harvested. Quantitative RT-PCR was performed using specific primers for MEIP (C), IP1 (D), or IP2 (E). Expression levels were normalized to GAPDH and compared to Mock (**p<0.003, ***p<0.0007, ****p<0.0001 [one-way ANOVA with Tukey’s multiple comparison test]).

Article Snippet: HEK293 and adult normal human dermal fibroblasts (NHDF) were obtained from ATCC and cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% heat-inactivated fetal bovine serum (FBS; Hyclone), 100 units/ml penicillin, 100 μg/ml streptomycin, and 100 μg/ml glutamine (Thermofisher).

Techniques: Infection, Staining, Software, Comparison, Quantitative RT-PCR, Expressing

Journal: Nanomaterials

Article Title: Chelidonium majus L. Incorporated Emulsion Electrospun PCL/PVA_PEC Nanofibrous Meshes for Antibacterial Wound Dressing Applications

doi: 10.3390/nano11071785

Figure Lengend Snippet: Evaluation of the NHDF cell after 1, 3, and 7 days in direct contact with the produced electrospun PCL/PVA_PEC nanofibrous meshes with and without C. majus extract. (“n.s.” indicates not significant ( p > 0.05)).

Article Snippet: Normal human dermal fibroblasts (NHDF) cells were acquired from ATCC (American Type Culture Collection, Manassas, VA, USA).

Techniques: Produced

Journal: International Journal of Molecular Sciences

Article Title: Functionalization of Bacterial Cellulose with the Antimicrobial Peptide KR-12 via Chimerical Cellulose-Binding Peptides

doi: 10.3390/ijms25031462

Figure Lengend Snippet: Solution peptide effect on NHDF viability assessed through metabolic activity. All data are normalized to the metabolic activity of untreated control cells indicated by the red dashed line; 70% activity is indicated by the grey dashed line. Data are presented as the mean and standard deviation from three independent experiments ( n = 3 per experiment). Statistical differences determined by an ANOVA followed by the Tukey’s HSD test (* p < 0.05, ** p < 0.01, *** p < 0.005).

Article Snippet: Komagataeibacter hansenii NQ5 ( K. hansenii ATCC 53582), normal human dermal fibroblasts (NHDFs; American Type Culture Collection: CRL-2565), Staphylococcus aureus (ATCC 43866, strain designation: LRA 44.01.83), Escherichia coli (ATCC 33694, strain designation: HB101) and Pseudomonas aeruginosa (ATCC 29260, strain designation: PA-103) were purchased from the American Type Culture Collection (ATCC), Manassas, VA, USA, while human endothelial keratinocyte cells (HaCaT, catalog number: T0020001) were purchased from AddexBio, San Diego, CA, USA.

Techniques: Activity Assay, Control, Standard Deviation

Journal: International Journal of Molecular Sciences

Article Title: Functionalization of Bacterial Cellulose with the Antimicrobial Peptide KR-12 via Chimerical Cellulose-Binding Peptides

doi: 10.3390/ijms25031462

Figure Lengend Snippet: Functionalized BC effect on NHDF and HaCaT viability. Normalized metabolic activity of ( A , B ) NHDF and ( C , D ) HaCaT after exposure to unfunctionalized BC (BC control) and BC functionalized with Long-CBP-KR12 and Short-CBP-KR12 normalized to untreated cells. Both the ( A , C ) opaque and ( B , D ) transparent BC were evaluated. All data are normalized to the metabolic activity of untreated control wells indicated by the red dashed line, and 70% activity is indicated by the grey dashed line. Data are presented as the mean and standard deviation from three independent experiments ( n = 3 per experiment). No statistical differences determined by ANOVA.

Article Snippet: Komagataeibacter hansenii NQ5 ( K. hansenii ATCC 53582), normal human dermal fibroblasts (NHDFs; American Type Culture Collection: CRL-2565), Staphylococcus aureus (ATCC 43866, strain designation: LRA 44.01.83), Escherichia coli (ATCC 33694, strain designation: HB101) and Pseudomonas aeruginosa (ATCC 29260, strain designation: PA-103) were purchased from the American Type Culture Collection (ATCC), Manassas, VA, USA, while human endothelial keratinocyte cells (HaCaT, catalog number: T0020001) were purchased from AddexBio, San Diego, CA, USA.

Techniques: Activity Assay, Control, Standard Deviation

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Resveratrol-Enriched Rice Attenuates UVB-ROS-Induced Skin Aging via Downregulation of Inflammatory Cascades

doi: 10.1155/2017/8379539

Figure Lengend Snippet: The levels of ROS in NHDFs treated as indicated for 24 h were measured using flow cytometry with DCFH-DA dye. The number of cells is plotted against the dichlorofluorescein fluorescence detected by the FL-1 channel (a). The relative ROS production of the cells is shown in each histogram (b). Values are mean ± SEM. The labels # and ∗ indicate significant differences ( p < 0.05) when compared with the normal control and UV (+) control, respectively. ### p < 0.001 versus the normal control, ∗∗ p < 0.01 versus the UVB-irradiated control. NC is normal control, Ctl is UVB control, NR is normal rice, RR is resveratrol-enriched rice, and R is resveratrol. NR and RR were treated in μ g/mL, and R was treated in μ M.

Article Snippet: Normal human dermal fibroblast cells (NHDFs) were obtained by skin biopsy from a healthy young male donor (MCTT Core Inc., Seoul, Korea).

Techniques: Flow Cytometry, Fluorescence, Control, Irradiation