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: Oncotarget

Article Title: Expression and potential role of cellular retinol binding protein I in psoriasis

doi: 10.18632/oncotarget.26314

Figure Lengend Snippet: (A-B) , bar graphs of NF-kB and AKT signaling-specific genes array of CRBPI-transfected HaCaT cells. (C) Representative blots and (D) bar graphs densitometric evaluation of pAKT/AKT, pErk, IL-6 and IL-8 protein expression. Significant changes reported * p < 0.05, ** p < 0.001; *** p < 0.0005 at Student’s t -test.

Article Snippet: Immortalized human keratinocyte cells HaCaT, (Lonza, Milano, Italy) maintained in D-MEM (Lonza Bio Pharma AG, Switzerland) were transfected by using a vector pTargeT Mammalian expression system carrying the whole sequence of CRBPI gene (NM_002899) and the gene for G418 resistance (Promega, Italy), or the G418-resistance gene alone.

Techniques: Transfection, Expressing