Journal: Pain

Article Title: MicroRNA-mediated GABA Aα-1 receptor subunit down-regulation in adult spinal cord following neonatal cystitis-induced chronic visceral pain in rats.

doi: 10.1016/j.pain.2012.09.002

Figure Lengend Snippet: Fig. 4. (A) Real-time RT-PCR analysis of differentially expressed GABAAa-1 gene in spinal dorsal horns (L6–S1) from rats with neonatal cystitis. Two different treatment protocols were followed. Tissues were examined on P60 from the unchallenged protocol and on P30 from the re-challenged protocol. The ordinate value 2DCT corresponds to the mRNA expression relative to reference gene ribosomal RNA (rRNA); data are represented as mean ± SD (n = 5 per group). (B) Western blot analysis of GABAAa-1 protein in spinal dorsal horn samples (L6–S1) from rats with neonatal cystitis. The intensity of GABAAa-1 immunoreactivity for different tissues was normalized against the intensity of b-actin expression for the same tissue. Results are expressed as means ± SD (n = 5 per group). ⁄Significant difference at p < 0.05. (C) GABAAa-1 immunostaining of differentially expressed GABAAa-1 gene in spinal dorsal horns (L6–S1) from rats with neonatal cystitis. The tissues were examined on P60 after zymosan/saline treatment at P14 to P16. The scale bar is 50 lm. The intensity of staining for 10 individual cells from each group was determined; data are presented as means ± SD. ⁄Significant difference at p < 0.001.

Article Snippet: GABAAa-1 gene expression in spinal dorsal horn neurons (L6–S1) by real-time quantitative RT-PCR Total RNA was extracted from spinal dorsal horn samples from both the unchallenged (P60) and re-challenged (P30) groups of animals using Total RNA extraction kit from Bio-Rad (#732-6830).

Techniques: Quantitative RT-PCR, Expressing, Western Blot, Immunostaining, Saline, Staining

Journal: Archives of dermatological research

Article Title: Control of late cornified envelope genes relevant to psoriasis risk: upregulation by 1,25-dihydroxyvitamin D3 and plant-derived delphinidin.

doi: 10.1007/s00403-013-1390-1

Figure Lengend Snippet: Fig. 3 Regulation of selected LCE genes in a human keratinocyte cell line, HaCaT. Cells were incubated for 24 h with ligands at a final concentration of 10 nM for 1,25D, 10 lM for docosahexaenoic acid (DHA), 5 lM for curcumin (CM) and 10 lM for delphinidin (Del). All ligands were dissolved in ethanol except for CM, which was dissolved in DMSO with a separate vehicle control. RNA isolation, first strand cDNA synthesis and real-time PCR were as described in ‘‘Materials and methods’’. The results shown are a composite of at least seven independent experiments. The high and low values for each treatment were discarded and averages of the remaining five values are shown as ±SEM with each sample assayed in triplicate. Single asterisks denote averages that are statistically significant from ethanol control, p \ 0.05; triple asterisks indicate p \ 0.001

Article Snippet: Real-time PCR analysis of LCE gene expression RNA was isolated using an Aurum Total RNA Mini Kit (BioRad, Hercules, CA, USA) from cells grown in 60 mm dishes to approximately 70 % confluence.

Techniques: Incubation, Concentration Assay, Control, Isolation, cDNA Synthesis, Real-time Polymerase Chain Reaction

Journal: Archives of dermatological research

Article Title: Control of late cornified envelope genes relevant to psoriasis risk: upregulation by 1,25-dihydroxyvitamin D3 and plant-derived delphinidin.

doi: 10.1007/s00403-013-1390-1

Figure Lengend Snippet: Fig. 4 Regulation of LCE genes by 1,25D in primary human keratinocytes. HEKn cells were plated at 550,000 cells per 60 mm plate, incubated overnight, then treated as follows: a 24 h with 1,25D (three concentrations) or ethanol vehicle without high calcium preincubation; b preincubation for 24 h with 1.2 mM calcium, then with 1,25D or ethanol vehicle for 24 h. RNA isolation, synthesis of first strand DNA and real-time PCR are described in ‘‘Materials and methods’’. Results are from three (b) or four (a) independent experiments, each in triplicate, ±SEM. Single asterisks denote averages that are statistically significant from ethanol control, p \ 0.05; double asterisks denote p \ 0.01; triple asterisks indicate p \ 0.001

Article Snippet: Real-time PCR analysis of LCE gene expression RNA was isolated using an Aurum Total RNA Mini Kit (BioRad, Hercules, CA, USA) from cells grown in 60 mm dishes to approximately 70 % confluence.

Techniques: Incubation, Isolation, Real-time Polymerase Chain Reaction, Control

Journal: Archives of dermatological research

Article Title: Control of late cornified envelope genes relevant to psoriasis risk: upregulation by 1,25-dihydroxyvitamin D3 and plant-derived delphinidin.

doi: 10.1007/s00403-013-1390-1

Figure Lengend Snippet: Fig. 5 Ability of delphinidin, a novel candidate VDR ligand, to regulate CYP24A1, LCE2B and all five genes in the LCE3 cluster. Cells were plated and dosed with or without 24 h of 1.2 mM calcium preincubation as described in the legend of Fig. 4, and RNA isolations, first strand cDNA synthesis and real-time PCR were performed as described in ‘‘Materials and methods’’. Results are from three independent experiments, each assayed in triplicate, ±SEM, except for results with 10 lM delphinidin, for which four independent experiments were carried out. Single asterisks denote averages that are statistically significant from ethanol control, p \ 0.05; double asterisks denote p \ 0.01

Article Snippet: Real-time PCR analysis of LCE gene expression RNA was isolated using an Aurum Total RNA Mini Kit (BioRad, Hercules, CA, USA) from cells grown in 60 mm dishes to approximately 70 % confluence.

Techniques: cDNA Synthesis, Real-time Polymerase Chain Reaction, Control