of the two peptides as products of NFIX mRNA. The last lane shows NFIX expression in HEK 293T cells, used in transient transfection experiments. Two lanes for each construct shown in,, and represent the expression from two independent clones for each construct. Found at: doi:10.1371/journal.pone.0005050.s002 C-terminal FLAG tagged peptide is primarily present out of nucleus. Rarely, it was seen strongly in nucleus also. N-terminal HA-tagged peptide is nuclear in most cells and infrequently present in cytoplasm too. NFIX antibody recognized endogenously expressed NFIX as present all over the cells. Staining with no primary antibody and pre-immune serum in case of NFIX antibody were used as negative controls. Found at: doi:10.1371/journal.pone.0005050.s003 Real time PCRs and qRTPCRs Total RNA was isolated and cDNA was synthesized using reverse transcription kit. ChIP DNA and input DNA were obtained as described above. Equal amounts of cDNA and equal volumes of ChIP DNA were used as template for each assay. GAPDH was used as internal control in all qRTPCRs and ACTB was used an additional internal control in some cases. All reactions were performed using 
SYBR-green premix and values calculated using delta-delta Ct method. Melt curve analysis. Values for serum negative MedChemExpress Tedizolid (phosphate) controls are not shown in any graph because the Ct values were too high or not obtained. All primer sequences are mentioned in reproducibility of NFIX-siRNA-induced changes in global gene expression. The hybridizations represented by transfection number 6 are the most different from the remaining 5 for most of the reporters. Found at: doi:10.1371/journal.pone.0005050.s005 Supporting Information Sequence alignment of prey clones identified in the yeast-2-hybrid screen with RefSeq RNA sequences. Two different clones of HMGN1 and one clone of CGGBP1. Stars indicate exact match. Found at: doi:10.1371/journal.pone.0005050.s006 The recent emergence of novel synthetic psychoactive drugs and their sale through internet sites has raised concerns about the potential harms associated with compounds which lack any formal toxicology profiles. Among the novel psychoactive substances that have emerged in recent years are methoxetamine -2-2-cyclohexanone), which is an analogue of ketamine -2–2-cyclohexanone), methoxetamine’s close deoxy-analogue 3-methoxyeticyclidine cyclohexanamine), and both the 3- and 4-methoxy analogues of phencyclidine, namely 1-piperidine and 1-piperidine. Methoxetamine, also known as `MXE’, `MXE-powder’, `METHO’, and `MEXXY’ has gained some prominence in the United Kingdom as a legal alternative to ketamine . Phencyclidine and the related compounds eticyclidine, rolicyclidine and tenocyclidine are controlled substances, but recently 3methoxy-PCP, 4-methoxy-PCP, and 3-methoxy-PCE have emerged as legally available alternatives to 16483784 PCP. Pharmacologically, ketamine’s main action is on glutamatergic transmission, the major excitatory neurotransmitter system in the 18772318 brain. It is a non-competitive antagonist at one of the three glutamate receptor subtypes, the N-methyl-D-aspartate receptor. The NMDA receptor is also considered to be a key pharmacological target for phencyclidine. Although there is little information available on the novel ketamine and PCP analogues, their behavioural effects in human subjects resemble those induced by ketamine and PCP, characteristic of dissociative anaesthetics. The wanted effects include euphoria, empathy, dissociation from the physical bod
