Journal: Molecular Pain
Article Title: Expression of macrophage migration inhibitory factor in footpad skin lesions with diabetic neuropathy
Figure Lengend Snippet: Effects of macrophage migration inhibitory factor (MIF) small interfering RNA (siRNA) in the presence of 400 μm methylglyoxal (MG) in human HaCaT keratinocytes. Cells were transfected with MIF siRNA or a control siRNA as well as recombinant human (rh)-MIF before stimulation with MG. Expression levels of MIF, glyoxalase I (GLO-I), and intraepidermal nerve fibers (IENF) mRNAs and proteins were measured by real-time reverse transcription polymerase chain reaction (a) to (c) and Western blot (d) to (g), respectively. Real-time PCR analysis was performed with a SYBR Green assay system. Relative gene expression levels normalized to β-actin was calculated with 2 −△△CT method. As a control for Western blot analysis, the level of β-actin was determined using an antibody against β-actin. All results are presented as mean ± SD of three independent experiments. ++ P
Article Snippet: Rabbit polyclonal antibody against MIF (1:200, Santa Cruz) and mouse monoclonal antibodies against IENF (1:200), GLO-I (1:1000) (Abcam, Cambridge, MA, USA) or β-actin (1:200, Santa Cruz) were used in this study.
Techniques: Migration, Small Interfering RNA, Transfection, Recombinant, Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Real-time Polymerase Chain Reaction, SYBR Green Assay