Journal: Gels

Article Title: Oleogel Dressings for Skin Therapy: Physicochemical and Bioactive Properties of Cosmetic Oil-Based Systems Enriched with Essential Oils

doi: 10.3390/gels12030248

Figure Lengend Snippet: Dose-dependent viability profile of HaCaT keratinocytes exposed to optimized oleogels formulations (SFW, RBW, HSA) over 24 and 48 h. Data are expressed as mean ± SD (n = 3). Different letters: significant difference between treatments ( p < 0.05). Concentration (μg/mL): essential oil blend equivalent.

Article Snippet: The viability of immortalized human keratinocytes (HaCaT, CLS 300493; Cell Lines Service GmbH, Germany) was assessed.

Techniques: Concentration Assay

Journal: PLoS ONE

Article Title: PharmDB-K: Integrated Bio-Pharmacological Network Database for Traditional Korean Medicine

doi: 10.1371/journal.pone.0142624

Figure Lengend Snippet: (A) Possible effect of caffeic acid in Scrophulariae Radix and Malvae Semen. (B) HaCaT cells were pretreated with caffeic acid (50 and 100 μM) for 1 hr, and then cells were exposed to TPA (100 nM) for additional 8 hr. (C) Cells were treated with TPA (100 nM) in the presence of caffeic acid (50 and 100 μM) for 2 hr. The NF-κB DNA binding activity was assessed by the gel-shift assay. The nuclear extracts were prepared and incubated with the radiolabeled oligonucleotides containing κB consensus sequence for the analysis of NF-κB DNA binding by EMSA. (D) Nuclear proteins were separated by 10% SDS-polyacrylamide gel electrophoresis and immunoblotted with p65 antibody. Lamin B was used as markers of nuclear proteins. (E) The cytosolic extracts prepared from cells incubated with TPA for 3 hr in the presence or absence of caffeic acid were immunoblotted with was analyzed by Western blotting to examine the expression of IκBα. (F) HaCaT cells were treated with TNF-α (20 nM) in the absence or presence of caffeic acid (100 μM) for 24 hr and then the isolated RNA was reverse-transcribed and amplified as described in Materials and Methods. Expression of il-8 and gapdh mRNA was measured by RT-PCR.

Article Snippet: HaCaT cells were kindly gifted from Dr. Zigang Dong (Hormel Institute, University of Minnesota, MN, USA) and were maintained routinely in DMEM medium supplemented with 10% fetal bovine serum and a 100 ng/ml penicillin/streptomycin/fungizone mixture at 37°C in a humidified atmosphere of 5% CO2/95% air.

Techniques: Binding Assay, Activity Assay, Gel Shift, Incubation, Sequencing, Polyacrylamide Gel Electrophoresis, Western Blot, Expressing, Isolation, Reverse Transcription, Amplification, Reverse Transcription Polymerase Chain Reaction

Journal: Scientific Reports

Article Title: Different signaling patterns contribute to loss of keratinocyte cohesion dependent on autoantibody profile in pemphigus

doi: 10.1038/s41598-017-03697-7

Figure Lengend Snippet: Structural changes in keratinocytes after 24 h of IgG treatment. HaCaT keratinocytes stably expressing CK5-YFP were exposed to different IgG fractions for 24 h. Images shown are representatives of four independent experiments. Dsg3 staining ( a – f ) and CK5 expression ( g – l ) after IgG treatment. ( m – r ) Merged images of Dsg3 staining and CK5-YFP. ( s – x ) CK5 fluorescence intensity of a 100 pixel long line placed centrally over the cell membrane (n = 4). ( y ) Analysis of the distance between fluorescence peaks shown in ( s – x ) in μ m (n = 4, *p ≤ 0.05 vs. c-IgG). Bar represents 10 μ m.

Article Snippet: The human immortalized keratinocyte cell line HaCaT was cultured in Dulbecco’s Modified Eagle Medium containing 10% FCS (Biochrom, Berlin, Germany), 50 U/ml penicillin and 50 g/ml streptomycin (both AppliChem, Darmstadt, Germany) in a humidified atmosphere of 5% CO 2 at 37 °C HaCaT cells stably transfected with keratin5-YFP were used for immunostaining experiments (kind gift of Reinhard Windoffer and Nicole Schwarz, Institute of Molecular and Cellular Anatomy, RWTH Aachen University ).

Techniques: Stable Transfection, Expressing, Staining, Fluorescence, Membrane

Journal: Scientific Reports

Article Title: Different signaling patterns contribute to loss of keratinocyte cohesion dependent on autoantibody profile in pemphigus

doi: 10.1038/s41598-017-03697-7

Figure Lengend Snippet: Signaling pathways activated by different IgG fractions. ( a ) Triton fractionation of HaCaT lysates into a TX-100 insoluble cytoskeletal and a TX-100 soluble non-cytoskeletal pool. Representative cropped Western blots of p38MAPK, src and Erk activity for each pool. ( b ) Densitometric analysis of signaling molecule phosphorylation normalized to the total amount in percent of control (n = 5–14, *p ≤ 0.05 vs. c-IgG). ( c ) Erk activation after inhibition of PKC by Bim-X and subsequent incubation of mc-PV and PF-IgG for 30 min. ( d ) Densitometric analysis of Erk phosphorylation corresponding to c (n = 4, *p ≤ 0.05 vs. respective c-IgG condition). Ratiometric fluorescent intensity measurement of Fura-2-AM at 340/380 nm normalized to baseline. ( e ) Graphs of c-IgG and PF-IgG to illustrate experimental setup. IgG incubation started at 20 s (arrow) and ( f ) concentrations were analyzed 60 s afterwards (dotted line) (n = 3, *p ≤ 0.05 vs. c-IgG). ( g ) Measurement of intracellular cAMP concentrations in keratinocytes after 6 h of IgG treatment (n = 4; *p ≤ 0.05 vs. c-IgG).

Article Snippet: The human immortalized keratinocyte cell line HaCaT was cultured in Dulbecco’s Modified Eagle Medium containing 10% FCS (Biochrom, Berlin, Germany), 50 U/ml penicillin and 50 g/ml streptomycin (both AppliChem, Darmstadt, Germany) in a humidified atmosphere of 5% CO 2 at 37 °C HaCaT cells stably transfected with keratin5-YFP were used for immunostaining experiments (kind gift of Reinhard Windoffer and Nicole Schwarz, Institute of Molecular and Cellular Anatomy, RWTH Aachen University ).

Techniques: Protein-Protein interactions, Fractionation, Western Blot, Activity Assay, Phospho-proteomics, Control, Activation Assay, Inhibition, Incubation

Journal: Scientific Reports

Article Title: Different signaling patterns contribute to loss of keratinocyte cohesion dependent on autoantibody profile in pemphigus

doi: 10.1038/s41598-017-03697-7

Figure Lengend Snippet: Adhesion measurements with modulation of signaling molecules. Dispase-based dissociation assays in HaCaT keratinocytes after applying the specific inhibitors SB202190 (p38MAPK), Bim-X (PKC), U0126 (Erk) or PP2 (Src) for 1 h, followed by either ( a ) 24 h incubation of IgG fractions or ( c ) 2 h incubation of IgG fractions (n = 4; *p ≤ 0.05 vs. c-IgG with the respective mediator). ( b , d ) Representative images of monolayers incubated with 10 μ m MTT to visualize cell fragments.

Article Snippet: The human immortalized keratinocyte cell line HaCaT was cultured in Dulbecco’s Modified Eagle Medium containing 10% FCS (Biochrom, Berlin, Germany), 50 U/ml penicillin and 50 g/ml streptomycin (both AppliChem, Darmstadt, Germany) in a humidified atmosphere of 5% CO 2 at 37 °C HaCaT cells stably transfected with keratin5-YFP were used for immunostaining experiments (kind gift of Reinhard Windoffer and Nicole Schwarz, Institute of Molecular and Cellular Anatomy, RWTH Aachen University ).

Techniques: Incubation