Journal: Frontiers in Cell and Developmental Biology

Article Title: Fisetin Promotes Hair Growth by Augmenting TERT Expression

doi: 10.3389/fcell.2020.566617

Figure Lengend Snippet: Screening for polyphenols that activate hTERT transcription in HaCaT cells. (A) Screening for polyphenols that activate hTERT transcription. Polyphenols (10 μM) were added to the HaCaT(hTERTp-EGFP) cells and cultured for 48 h; then, changes in EGFP fluorescence were monitored using an IN Cell Analyzer 1000. (B) The effect of polyphenols on the expression of endogenous hTERT in HaCaT cells was evaluated by qRT-PCR. Statistical significance was determined using a two-sided Student’s t -test. Statistical significance was defined as P < 0.05 (** P < 0.01; *** P < 0.001).

Article Snippet: The HaCaT human keratinocyte cell line (Riken Bioresource Center, Tsukuba, Japan) was cultured in Dulbecco’s Modified Eagle’s Medium (DMEM; Nissui, Tokyo, Japan) supplemented with 10% fetal bovine serum (FBS; Life Technologies, Gaithersburg, MD, United States) at 37°C in a 5% CO 2 atmosphere.

Techniques: Cell Culture, Fluorescence, Expressing, Quantitative RT-PCR

Journal: Frontiers in Cell and Developmental Biology

Article Title: Fisetin Promotes Hair Growth by Augmenting TERT Expression

doi: 10.3389/fcell.2020.566617

Figure Lengend Snippet: Effects of polyphenols on the expression of cytokine-encoding genes in HaCaT cells. After HaCaT cells were treated with polyphenols, the gene expression levels of IGF-1 (A) , KGF (B) , and TGF- β 1 (C) were measured by qRT-PCR. Statistical significance was determined using a two-sided Student’s t -test. Statistical significance was defined as P < 0.05 (* P < 0.05; ** P < 0.01; *** P < 0.001).

Article Snippet: The HaCaT human keratinocyte cell line (Riken Bioresource Center, Tsukuba, Japan) was cultured in Dulbecco’s Modified Eagle’s Medium (DMEM; Nissui, Tokyo, Japan) supplemented with 10% fetal bovine serum (FBS; Life Technologies, Gaithersburg, MD, United States) at 37°C in a 5% CO 2 atmosphere.

Techniques: Expressing, Gene Expression, Quantitative RT-PCR

Journal: Frontiers in Cell and Developmental Biology

Article Title: Fisetin Promotes Hair Growth by Augmenting TERT Expression

doi: 10.3389/fcell.2020.566617

Figure Lengend Snippet: Effects of polyphenols on β-catenin activity. The effect of these polyphenols on the activity of β-catenin was evaluated. (A) The TOP-Flash reporter assay was performed to evaluate β-catenin activity in HaCaT cells treated with polyphenols. (B) The effect of polyphenols on AXIN2 expression was evaluated by qRT-PCR. (C,D) The effect of polyphenols on protein expression of β-catenin was evaluated by western blotting using anti-β-catenin antibody. Band intensities were quantitatively determined using ImageJ software. (E) The effect of polyphenols on activity of β-catenin was evaluated by immunofluorescence study using anti-active β-catenin antibody. Statistical significance was determined using a two-sided Student’s t -test. Statistical significance was defined as P < 0.05 (* P < 0.05; ** P < 0.01; *** P < 0.001).

Article Snippet: The HaCaT human keratinocyte cell line (Riken Bioresource Center, Tsukuba, Japan) was cultured in Dulbecco’s Modified Eagle’s Medium (DMEM; Nissui, Tokyo, Japan) supplemented with 10% fetal bovine serum (FBS; Life Technologies, Gaithersburg, MD, United States) at 37°C in a 5% CO 2 atmosphere.

Techniques: Activity Assay, Reporter Assay, Expressing, Quantitative RT-PCR, Western Blot, Software, Immunofluorescence

Journal: Frontiers in Cell and Developmental Biology

Article Title: Fisetin Promotes Hair Growth by Augmenting TERT Expression

doi: 10.3389/fcell.2020.566617

Figure Lengend Snippet: Effect of polyphenols on the growth of HaCaT cells. (A) After HaCaT cells were treated with 10 μM of polyphenols, cell proliferation was monitored using the Cell Counting Kit-8. (B) The proliferation index on the 3rd day was determined. Statistical significance was determined using a two-sided Student’s t -test. Statistical significance was defined as P < 0.05 (* P < 0.05).

Article Snippet: The HaCaT human keratinocyte cell line (Riken Bioresource Center, Tsukuba, Japan) was cultured in Dulbecco’s Modified Eagle’s Medium (DMEM; Nissui, Tokyo, Japan) supplemented with 10% fetal bovine serum (FBS; Life Technologies, Gaithersburg, MD, United States) at 37°C in a 5% CO 2 atmosphere.

Techniques: Cell Counting

Journal: Frontiers in Cell and Developmental Biology

Article Title: Fisetin Promotes Hair Growth by Augmenting TERT Expression

doi: 10.3389/fcell.2020.566617

Figure Lengend Snippet: Role of hTERT in the polyphenol-induced effects on HaCaT cells. (A) Relative expression levels of hTERT in HaCaT cells transduced with retroviruses expressing shRNA targeting hTERT (sh-hTERT1 and 2) and scramble shRNA (SCR) were evaluated by qRT-PCR. HaCaT cells whose hTERT expression was reduced by shRNA (sh-hTERT-1 and 2) were treated with polyphenols, and relative β-catenin activity (B) , cytokine gene expression (C,D) , and cell proliferation (E) was determined. Statistical significance was determined using a two-sided Student’s t -test. Statistical significance was defined as P < 0.05 (* P < 0.05; ** P < 0.01; *** P < 0.001).

Article Snippet: The HaCaT human keratinocyte cell line (Riken Bioresource Center, Tsukuba, Japan) was cultured in Dulbecco’s Modified Eagle’s Medium (DMEM; Nissui, Tokyo, Japan) supplemented with 10% fetal bovine serum (FBS; Life Technologies, Gaithersburg, MD, United States) at 37°C in a 5% CO 2 atmosphere.

Techniques: Expressing, Transduction, shRNA, Quantitative RT-PCR, Activity Assay, Gene Expression

Journal: bioRxiv

Article Title: RPRM negatively regulates ATM levels involving its phosphorylation mediated by CDK4/CDK6

doi: 10.1101/2021.11.10.468148

Figure Lengend Snippet: a, b Co-immunoprecipitation assay (Co-IP) was performed to demonstrate the interaction between ATM and RPRM, which was enhanced in irradiated H460-RPRM cells. c Representative immunofluorescence images of co-localization of RPRM and ATM in RPRM -/- MEFs reexpressing RPRM at different times post 20 Gy X-irradiation. d, e Change in the levels of total ATM and phospho-ATM in H460-NC (N)/RPRM (R) cells at different times post 2 Gy X-irradiation. Quantification of ATM levels ( e ), which were the average of three independent experiments. f Change in the ATM levels of HaCaT-NC (N)/RPRM (R) cells irradiated with 2 Gy X-rays showed that RPRM overexpression in HaCaT cells decreased ATM levels upon IR. g, h The ATM levels of RPRM +/+ MEFs were lower than those of RPRM -/- MEFs at different times post 20 Gy X-irradiation. Bar graph ( h ) shows quantification of ATM levels. i, j RPRM reexpression reduced the ATM levels in RPRM -/- MEFs post 20 Gy X-irradiation. Bar graph ( j ) shows quantification of ATM levels. Experiments were independently repeated three times with similar results. * P < 0.05; ** P < 0.01.

Article Snippet: HaCaT human immortalized keratinocytes were obtained from China Center for Type Culture Collection.

Techniques: Co-Immunoprecipitation Assay, Irradiation, Immunofluorescence, Over Expression

Journal: PLOS ONE

Article Title: Aromatic oil from lavender as an atopic dermatitis suppressant

doi: 10.1371/journal.pone.0296408

Figure Lengend Snippet: Human keratinocyte-like cell line HaCaT stably expressing the NanoLuc luciferase reporter gene under the control of an XRE (XRE-NLuc::HaCaT) was incubated in culture medium containing 0.5 μM FICZ with or without the indicated amounts of agents for 24 h. The percentage inhibition of AD was calculated by the reporter expression with FICZ alone as 0%, indicating no inhibition, and without FICZ as 100% indicating complete inhibition. The dot markers in the graph represent data from three independent experiments. The vertical and horizontal bars represent the standard deviation and the mean, respectively. a) Comparison of L . angustifolia essential oil and generic lavender oil. b) Comparison of linalyl acetate, linalool, THL, and DHL. The statistical significance of the data points among the different concentrations and the dose-dependency were determined by the Kruskal-Wallis test or one-way ANOVA, and the Williams test, respectively, respectively. *, ** P <0.05 (n = 3).

Article Snippet: Human immortalized keratinocyte cell line HaCaT (#300493, CLI, Cosmo-bio, Tokyo, Japan), established from adult male skin, was maintained at 37°C under 5%CO 2 in Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with 10% fetal bovine serum (FBS).

Techniques: Stable Transfection, Expressing, Luciferase, Control, Incubation, Inhibition, Standard Deviation, Comparison

Journal: PLOS ONE

Article Title: Aromatic oil from lavender as an atopic dermatitis suppressant

doi: 10.1371/journal.pone.0296408

Figure Lengend Snippet: HaCaT cells were incubated in culture medium containing 5 μM FICZ with or without the indicated amounts of agents for 24 h. Total RNA was then extracted and reverse transcribed to cDNA that was subjected to qPCR using the primer set listed in S2 Table in . Artemin expression was normalized to GAPDH expression. Percentage inhibition was then calculated with FICZ alone as 0%, indicating no inhibition, and without FICZ as 100% indicating complete inhibition. The dot markers in the graphs represent data from two or three independent experiments. The vertical and horizontal bars represent the standard deviation and the mean, respectively. Comparisons of (a) L . angustifolia essential oil and generic lavender oil, and (b) linalyl acetate and linalool for inhibition of induced Artemin expression are shown. The statistical significance of the data points among the different concentrations and the dose-dependency were determined by the Kruskal-Wallis test or one-way ANOVA, and the Williams test, respectively. *, ** P <0.05 (n = 3).

Article Snippet: Human immortalized keratinocyte cell line HaCaT (#300493, CLI, Cosmo-bio, Tokyo, Japan), established from adult male skin, was maintained at 37°C under 5%CO 2 in Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with 10% fetal bovine serum (FBS).

Techniques: Incubation, Reverse Transcription, Expressing, Inhibition, Standard Deviation

Journal: PLOS ONE

Article Title: Aromatic oil from lavender as an atopic dermatitis suppressant

doi: 10.1371/journal.pone.0296408

Figure Lengend Snippet: a, b) HaCaT cells incubated in culture medium containing the indicated agents for 24 h. Total RNA was then extracted and reverse transcribed to cDNA that was subjected to qPCR using the primer set listed in S2 Table in . AhR and ARNT expression was normalized to GAPDH expression. Fold induction was calculated as the vehicle set at 1. Values in the graphs represent the mean ± SD of two independent experiments. There were no statistically significant differences under all conditions. c, d) HaCaT cells were incubated in culture medium containing the indicated agents for 24 h. c) Cells were lysed and subjected to western blotting with antibodies against AhR, ARNT, and α-tubulin. All the Western blots are shown from two (anti-ARNT) or three (anti-AhR and anti-gamma tubulin) independent experiments. The right-side numbers are experimental numbers. d) Band intensity of each lane was quantified by ImageJ and normalized to α-tubulin. The intensities are relative to the vehicle set at 1. Values in the graphs represent the mean ± SD of two or three independent experiments. * p <0.05, compared with FICZ alone. LA, linalyl acetate.

Article Snippet: Human immortalized keratinocyte cell line HaCaT (#300493, CLI, Cosmo-bio, Tokyo, Japan), established from adult male skin, was maintained at 37°C under 5%CO 2 in Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with 10% fetal bovine serum (FBS).

Techniques: Incubation, Reverse Transcription, Expressing, Western Blot

Journal: PLOS ONE

Article Title: Aromatic oil from lavender as an atopic dermatitis suppressant

doi: 10.1371/journal.pone.0296408

Figure Lengend Snippet: EC 1.5 and CV 70 in the keratinocyte activation assay.

Article Snippet: Human immortalized keratinocyte cell line HaCaT (#300493, CLI, Cosmo-bio, Tokyo, Japan), established from adult male skin, was maintained at 37°C under 5%CO 2 in Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with 10% fetal bovine serum (FBS).

Techniques: Activation Assay

Journal: PLOS ONE

Article Title: Aromatic oil from lavender as an atopic dermatitis suppressant

doi: 10.1371/journal.pone.0296408

Figure Lengend Snippet: Four key events in the skin sensitization reaction and reactivities of L . angustifolia essential oil, generic lavender oil, linalyl acetate, and linalool.

Article Snippet: Human immortalized keratinocyte cell line HaCaT (#300493, CLI, Cosmo-bio, Tokyo, Japan), established from adult male skin, was maintained at 37°C under 5%CO 2 in Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with 10% fetal bovine serum (FBS).

Techniques: Activation Assay, Lymph Node Assay

Journal: International Journal of Molecular Sciences

Article Title: Jellyfish Polysaccharides for Wound Healing Applications

doi: 10.3390/ijms231911491

Figure Lengend Snippet: Scratch test on HaCat human keratinocytes cell monolayers treated with RP-JSP1 and RP-JSP2. Control consists in untreated cells. ( a ) Representative micrographs (4× magnification) of treated and control monolayers; ( b ) confluency rate calculated as percentage of scratch closure in respect to the initial scratch area.

Article Snippet: Fibroblast BALB/3T3 clone A31 (CCL-163) and keratinocyte HaCat cell line were purchased from American Type Colture Collection (USA) and CellSystems (GmbH Germany), respectively.

Techniques: Control

Journal: Molecules

Article Title: Synergetic Effects of Aloe Vera Extract with Trimethylglycine for Targeted Aquaporin 3 Regulation and Long-Term Skin Hydration

doi: 10.3390/molecules29071540

Figure Lengend Snippet: Skin hydration after a single application of the formulas with and without the novel combination.

Article Snippet: The human keratinocyte cell line HaCaT was purchased from MatTek (MatTek Europe, Bratislava, Slovakia).

Techniques: Formulation, Negative Control

Journal: Molecules

Article Title: Synergetic Effects of Aloe Vera Extract with Trimethylglycine for Targeted Aquaporin 3 Regulation and Long-Term Skin Hydration

doi: 10.3390/molecules29071540

Figure Lengend Snippet: Skin hydration after long-term use of the formulas with and without the novel combination.

Article Snippet: The human keratinocyte cell line HaCaT was purchased from MatTek (MatTek Europe, Bratislava, Slovakia).

Techniques: Formulation, Negative Control