Journal: PLoS Pathogens

Article Title: Long non-coding RNA KIKAT/LINC01061 as a novel epigenetic regulator that relocates KDM4A on chromatin and modulates viral reactivation

doi: 10.1371/journal.ppat.1009670

Figure Lengend Snippet: (A and B) Immunoblotting (A) and RT-qPCR (B) analysis of the expression of K-bZIP, Orf45, Orf57, K8.1 in iSLK-BAC16 cells treated with 1 μg/ml Dox for 0, 24, 48 and 72 hours. (C) KSHV virion-associated DNA was determined by TaqMan qPCR. (D) Expression of KIKAT/LINC01061 was determined by RT-qPCR. (E) Immunoblotting of K-Rta in iSLK cells treated with or without Dox for 48 hours. (F) RT-qPCR analysis of KIKAT/LINC01061 in iSLK cells treated as described in (E). (G) The luciferase reporter plasmid containing KIKAT/LINC01061 promoter (TSS ± 500 bp) was co-transfected with pcDNA3-HA-K-Rta or with empty vector into 293T cells. After 48 hours, cells were collected for immunoblotting with anti-HA and anti-α-Tubulin antibodies. (H) Luciferase reporter assays were performed in 293T cells treated as described in (G). The activity of firefly luciferase was normalized to that of Renilla luciferase in the same assayed sample and reported as the relative activity of vector control. (I) iSLK-BAC16 cells were transfected with siKIKAT/LINC01061 or with control siGLO. After 6 hours, cells were re-seeded in 6-well plates and treated with 1 μg/ml Dox for another 48 hours. Relative expression of KIKAT/LINC01061 (upper panel) and KSHV virion-associated DNA (lower panel) was determined by RT-qPCR and TaqMan qPCR, respectively. (J) iSLK-BAC16 cells were transduced with lentivirus expressing full-length KIKAT/LINC01061 or with no cDNA (Mock). After 16 hours, cells were re-seeded in 6-well plates and treated with 1 μg/ml Dox for another 72 hours. Relative expression of KIKAT/LINC01061 (upper panel) and KSHV virion-associated DNA (lower panel) was determined as described in (I). (K) iSLK-BAC16 cells were transduced with lentivirus expressing full-length KIKAT/LINC01061 or with no cDNA (Mock). After 72 hours, relative expression of KIKAT/LINC01061 (upper panel) and KSHV virion-associated DNA (lower panel) was determined as described in (I).

Article Snippet: Primary antibodies against KDM4A (Polyclonal antibody purified from rabbit) [ ], K-bZIP (1:1000; Santa Cruz Biotechnology, sc-69797), Orf45 (1:1000; Santa Cruz Biotechnology, sc-53883), HA tag (1:4000; Cell Signaling Technology, #3724) and α-Tubulin (1:4000; Sigma, T6074-200UL) were used in this study.

Techniques: Western Blot, Quantitative RT-PCR, Expressing, Luciferase, Plasmid Preparation, Transfection, Activity Assay, Control, Transduction

Journal: eneuro

Article Title: Behavioral Comorbidities and Drug Treatments in a Zebrafish scn1lab Model of Dravet Syndrome

doi: 10.1523/eneuro.0066-17.2017

Figure Lengend Snippet: Figure 4. Treatment with antiepileptic drugs at 250 M. Statistically significant decreases in locomotion (distance traveled) and duration in the center of the open field were observed after treatment of zebrafish larvae with 250 M diazepam (C, D) or clemizole (G, H), but not after treatment with valproic acid (A, B) or trazodone (E, F; N 18 per group). Data are plotted in 30-s time bins showing mean SEM (left), and the 5-min total for each all individuals, are plotted on the right. p 0.001.

Article Snippet: Valproic acid, a broad spectrum antiepileptic drug (Tomson et al., 2016) commonly used in DS (Chiron and Dulac, 2011), exerts protective effects in larval or adult zebrafish exposed to the chemoconvulsant pentylenetetrazole: (1) decreasing behavioral or electrographic seizure activity and (2) improv- ing deficits in learning of a passive avoidance response (Lee et al., 2010).

Techniques: