Journal: Molecular Cancer Therapeutics

Article Title: Cell Intrinsic Role of COX-2 in Pancreatic Cancer Development

doi: 10.1158/1535-7163.mct-12-0342

Figure Lengend Snippet: Figure 7. Decreased FKBP5 negative feedback leads to enhanced AKT activation in Pdx1-Creþ;K-rasG12D/þ;Ptenlox/þ; Cox-2lox/lox mice. A, FKBP5 mRNA expression in PDACs of Pdx1-Creþ; K-rasG12D/þ;Ptenlox/þ;Cox-2lox/lox

Article Snippet: The following primary antibodies were used: phospho-AKT(Ser473) (Cell Signaling; 1:50), Cytokeratin 19 (ab15463, Abcam; 1:100), COX-2 (SP21; Thermo Scientific, ready-to-use),GRP78(11587-1AP,ProteinTechGroup; 1:50), and FKBP5 (14155-1-AP, ProteinTech Group, 1:50).

Techniques: Activation Assay, Expressing

Journal: Frontiers in Neuroscience

Article Title: NPY1 Receptor Agonist Modulates Development of Depressive-Like Behavior and Gene Expression in Hypothalamus in SPS Rodent PTSD Model

doi: 10.3389/fnins.2017.00203

Figure Lengend Snippet: Changes in mRNA levels for several genes in the mediobasal hypothalamus . Data are presented as mean ± SEM. n = 8–10 per group. (A) CRF mRNA levels. *** p < 0.001 compared to controls; ## p < 0.01 compared to SPS/V. (B) GR mRNA levels. ** p < 0.01, *** p < 0.001 compared to controls; # p < 0.05, ### p < 0.001 compared to SPS/V; &&& p < 0.001 compared to SPS/LeuPro. (C) FKBP5 mRNA levels. *** p < 0.001 compared to Controls; # p < 0.05 ### p < 0.001 compared to SPS/V; &&& p < 0.001 compared to SPS/LeuPro.

Article Snippet: For qPCR, the cDNA (33.2 ng in 2 μl) was mixed with 12.5 μl of FastStart Universal SYBR Green Master Rox (Roche Diagnostics, Indianapolis, IN) and 1 μl of the following primer sets: CRF (Crh, NM_031019.1, cat. no. PPR44803B, Qiagen); glucocorticoid receptor (GR) (Nt3c1, NM_012576.2, cat no. PRR52805B, Qiagen); FKBP5 (Fkbp5, NM_001012174.1, cat. no. PPR51629B, Qiagen) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH; Gapdh, NM_017008.4, forward 5′-TGGACCACCCAGCCCAGCAAG-3′, reverse 5′-GGCCCCTCCTGTTGTTATGGGGT-3′), to a final volume 25 μl in PCR-96-Microplate (Axygen Scientific, Union City, CA).

Techniques:

Journal: Steroids

Article Title: The Requirement for p42/p44 MAPK Activity in Progesterone Receptor-Mediated Gene Regulation is Target Gene-Specific

doi: 10.1016/j.steroids.2012.12.014

Figure Lengend Snippet: A) T47D cells were cultured in serum-free medium for 24 hours before pretreatment with DMSO or U0126 (10 μM) for 1 hour, followed by treatment with ethanol or R5020 (10 nM) for 6 hours. PR and tubulin protein expression levels were determined by Western blot analysis. A representative experiment is shown. U0126 does not affect PR expression levels in the absence or presence of R5020. B) In parallel to the experiments in panel A, relative SGK and CCND1 mRNA expression was determined using real-time qPCR and normalized to 18S rRNA expression. SGK gene induction was insensitive to U0126, while CCND1 basal and hormone-mediated gene expression was decreased with U0126. * p<0.05 compared to respective treatment in DMSO group. C) T47D cells were cultured and pretreated as in panel A, followed by treatment with ethanol or R5020 (10 nM) for 24 hours. Relative SGK, NDRG1 and FKBP5 mRNA expression was determined using real-time qPCR and normalized to 18S rRNA expression. SGK gene induction was abrogated by U0126, while NDRG1 gene induction was robustly decreased by U0126. In contrast, FKBP5 gene induction was relatively insensitive to U0126. * p<0.05 compared to DMSO R5020. ****p<0.0001 compared to DMSO R5020.

Article Snippet: Primers for 18S (previously described in [ 30 ]), SGK and cyclin D1 (CCND1) (previously described in [ 20 ]), FK506 binding protein 5 (FKBP5) (sense 5′-GGATATACGCCAACATGTTCAA-3′, antisense 5′-CCATTGCTTTATTGGCCTCT-3′), N-myc downstream regulated 1 (NDRG1) (sense 5′-CTACCATGACATCGGCATGAA-3′, antisense 5′-CGTGGCAGACGGCAAAGT-3′), and interleukin 1 receptor, type 1 (IL1R1) (sense 5′-AATTGATGAAGATGACCCAGTGC-3′, antisense 5′-GCAGGATTTTCCACACTGTAATAGTCT-3′) were purchased from Sigma-Genosys (The Woodlands, TX).

Techniques: Cell Culture, Expressing, Western Blot