Journal: RNA Biology

Article Title: Altered Levels of Long NcRNAs Meg3 and Neat1 in Cell And Animal Models Of Huntington’s Disease

doi: 10.1080/15476286.2018.1534524

Figure Lengend Snippet: Summary of the levels of ncRNAs in different models of HD.

Article Snippet: SiRNA mediated knockdown of Neat1 and Meg3 siRNAs against mouse NEAT1 (FlexiTube GeneSolution GS66961 for Neat1, Cat no: LPM15183A) and MEG3 (FlexiTube GeneSolution GS17263 for Meg3, Cat No: GS17263) were purchased from Qiagen and used, following the protocol described by the manufacturer. siRNAs were used at a final concentration of 40nM.

Techniques: Transfection

Journal: RNA Biology

Article Title: Altered Levels of Long NcRNAs Meg3 and Neat1 in Cell And Animal Models Of Huntington’s Disease

doi: 10.1080/15476286.2018.1534524

Figure Lengend Snippet: Summary of protein and miRNA interactions of ncRNAs Meg3, Neat1, and Xist.

Article Snippet: SiRNA mediated knockdown of Neat1 and Meg3 siRNAs against mouse NEAT1 (FlexiTube GeneSolution GS66961 for Neat1, Cat no: LPM15183A) and MEG3 (FlexiTube GeneSolution GS17263 for Meg3, Cat No: GS17263) were purchased from Qiagen and used, following the protocol described by the manufacturer. siRNAs were used at a final concentration of 40nM.

Techniques:

Journal: RNA Biology

Article Title: Altered Levels of Long NcRNAs Meg3 and Neat1 in Cell And Animal Models Of Huntington’s Disease

doi: 10.1080/15476286.2018.1534524

Figure Lengend Snippet: Expression of genes in HD associated with Huntington’s disease pathway (KEGG: 05016 and PANTHER: {"type":"entrez-protein","attrs":{"text":"P00029","term_id":"124076962","term_text":"P00029"}} P00029 ) and coded for protein interacting partners of NEAT1 or MEG3.

Article Snippet: SiRNA mediated knockdown of Neat1 and Meg3 siRNAs against mouse NEAT1 (FlexiTube GeneSolution GS66961 for Neat1, Cat no: LPM15183A) and MEG3 (FlexiTube GeneSolution GS17263 for Meg3, Cat No: GS17263) were purchased from Qiagen and used, following the protocol described by the manufacturer. siRNAs were used at a final concentration of 40nM.

Techniques: Expressing

Journal: RNA Biology

Article Title: Altered Levels of Long NcRNAs Meg3 and Neat1 in Cell And Animal Models Of Huntington’s Disease

doi: 10.1080/15476286.2018.1534524

Figure Lengend Snippet: Summary of co-expressed genes of MEG3, NEAT1, and XIST.

Article Snippet: SiRNA mediated knockdown of Neat1 and Meg3 siRNAs against mouse NEAT1 (FlexiTube GeneSolution GS66961 for Neat1, Cat no: LPM15183A) and MEG3 (FlexiTube GeneSolution GS17263 for Meg3, Cat No: GS17263) were purchased from Qiagen and used, following the protocol described by the manufacturer. siRNAs were used at a final concentration of 40nM.

Techniques:

Journal: RNA Biology

Article Title: Altered Levels of Long NcRNAs Meg3 and Neat1 in Cell And Animal Models Of Huntington’s Disease

doi: 10.1080/15476286.2018.1534524

Figure Lengend Snippet: MEG3, NEAT1 and XIST interacting protein enriched with HD pathway.

Article Snippet: SiRNA mediated knockdown of Neat1 and Meg3 siRNAs against mouse NEAT1 (FlexiTube GeneSolution GS66961 for Neat1, Cat no: LPM15183A) and MEG3 (FlexiTube GeneSolution GS17263 for Meg3, Cat No: GS17263) were purchased from Qiagen and used, following the protocol described by the manufacturer. siRNAs were used at a final concentration of 40nM.

Techniques:

Journal: RNA Biology

Article Title: Altered Levels of Long NcRNAs Meg3 and Neat1 in Cell And Animal Models Of Huntington’s Disease

doi: 10.1080/15476286.2018.1534524

Figure Lengend Snippet: Transcription factors that bind within 5 Kb upstream sequences of NEAT1, MEG3, and XIST.

Article Snippet: SiRNA mediated knockdown of Neat1 and Meg3 siRNAs against mouse NEAT1 (FlexiTube GeneSolution GS66961 for Neat1, Cat no: LPM15183A) and MEG3 (FlexiTube GeneSolution GS17263 for Meg3, Cat No: GS17263) were purchased from Qiagen and used, following the protocol described by the manufacturer. siRNAs were used at a final concentration of 40nM.

Techniques: Sequencing

Journal: Nature Communications

Article Title: Ligand-specific endocytic dwell times control functional selectivity of the cannabinoid receptor 1

doi: 10.1038/ncomms5589

Figure Lengend Snippet: ( a ) HEK293 cells expressing SEP-CB1Rs were exposed to 10μM CP55940, ORG27569, WIN 55212-2 and 2-AG for 5, 10 and 15 min. Cell lysates were analysed using western blots with phospho-ERK1/2 (p-ERK1/2) or total ERK1/2. ( b ) Quantified time course in a showing ERK1/2 phosphorylation levels induced by 10 μM of each compound. ( c ) HEK293 cells expressing SEP-CB1Rs were exposed to 10μM CP55940, ORG27569, WIN 55212-2 and 2-AG for 5, 10 and 15 min with PTX pre-treatment, respectively. ( d ) Quantified time course in c showing ERK1/2 phosphorylation levels from cells pretreated with PTX. ( e , g , i , k ) HEK293 cells co-expressing SEP-CB1R and either control ( e ), β-arrestin-1 ( g ), β-arrestin-2 ( i ) siRNAs or β-arrestin-2 siRNA with PTX pretreatment ( k ) were exposed to 10 μM of CP55940, ORG27569, WIN 55212-2 and 2-AG, respectively, as indicated. ( f , h , j , l ). Graphs in f , h , j , l provide the quantified ERK1/2 phosphorylation levels induced by 10 μM of each compound as shown in e , g , i , k . Data represent the mean±s.e.m. of at least three independent experiments.

Article Snippet: Briefly, HEK293 cells that were 40–50% confluent in a six-well plate were transfected with the plasmid encoding rat CB1 and 2.6 mg of siRNA (Qiagen) sequences targeting β-arrestin 1, β-arrestin 2 or non-silencing RNA duplex for control.

Techniques: Expressing, Western Blot, Phospho-proteomics, Control

Journal: Nature Communications

Article Title: Ligand-specific endocytic dwell times control functional selectivity of the cannabinoid receptor 1

doi: 10.1038/ncomms5589

Figure Lengend Snippet: ( a ) HEK293 cells expressing SEP-CB1R were preincubated with 30μM dyngo-4a and imaged under TIRF before and after bath application of 10μM WIN55212-2. Representative kymograph shows prolonged SEP-CB1R dwell times in the presence of dyngo-4a. ( b ) HEK293 cells expressing SEP-CB1R were pre-incubated with 30μM dyngo-4a before exposure to agonists and compared with no dyngo-4a treatment. ( c ) Graph provides the quantified ERK1/2 phosphorylation induced by 10 μM of each compound shown in b . Data are expressed as a percentage of the level of phosphorylation at 5 min for each compound without dyngo-4a pre-treatment. ( d ) HEK293 cells expressing SEP-CB1Rs with and without dyngo-4a pretreatment were exposed to 10μM 2-AG as indicated with PTX pre-treatment. ( e ) HEK293 cells co-expressing SEP-CB1R and β-arrestin-1 siRNAs with and without dyngo-4a pretreatment were exposed to 10μM 2-AG as indicated. ( f ) Quantified time course in d , e showing ERK1/2 phosphorylation levels induced by 10μM 2-AG for 5, 15, 30, 45 and 60 min for cells either pre-treated with PTX or co-transfected with β-arrestin-1 siRNA. ( g ) HEK293 cells expressing SEP-CB1R were treated with PTX (or no PTX as a control), and then incubated with 30μM dyngo-4a before subsequent exposure to WIN 55,212-2. ( h ) Graph provides the quantified ERK1/2 phosphorylation induced by 10μM WIN 55,212-2 in the absence of PTX treatment shown in g . ( i ) HEK293 cells expressing SEP-CB1R with or without dominant-negative dynamin 2 K44A were exposed to agonists. ( j ) Graph provides the quantified ERK1/2 phosphorylation induced by 10 μM of each compound shown in i . All data are expressed as a percentage of the level of phosphorylation at 5 min for 2-AG without dyngo-4a pre-treatment and represent the mean±s.e.m. of at least three independent experiments.

Article Snippet: Briefly, HEK293 cells that were 40–50% confluent in a six-well plate were transfected with the plasmid encoding rat CB1 and 2.6 mg of siRNA (Qiagen) sequences targeting β-arrestin 1, β-arrestin 2 or non-silencing RNA duplex for control.

Techniques: Expressing, Incubation, Phospho-proteomics, Transfection, Control, Dominant Negative Mutation