Journal: Journal of Cellular and Molecular Medicine

Article Title: YAP accelerates vascular senescence via blocking autophagic flux and activating mTOR

doi: 10.1111/jcmm.15902

Figure Lengend Snippet: Knockdown of YAP inhibits cellular and vascular senescence. A, SA‐β‐gal staining was used to analyse senescence degree in HUVECs transfected with YAP siRNA (siYAP#1, siYAP#2, siYAP#3) or negative control siRNA (non‐targeting 20‐25 nt siRNA) for 6 h. B, SA‐β‐gal‐positive cell percentage. C, Western blot was preformed to analyse PYAP, YAP, P16, P21 and P53 in cells transfected with YAP siRNA (siYAP#1, siYAP#2, siYAP#3) or negative control siRNA for 6 h. D‐G, The semi‐quantification of the proteins in panel C respectively. H, Western blot was used to analyse YAP and senescence markers P16, P21 and P53 in vascular tissues treated with YAP siRNA (siYAP#1, siYAP#2, siYAP#3) or negative control siRNA for 6 h. I‐J, The semi‐quantification of the proteins in panel H, respectively. K, Expression of indicated proteins was analysed by immunoblot in nuclear and cytoplasmic protein extractions from in HUVECs treated with or without YAP siRNA (siYAP#1, siYAP#2, siYAP#3) or negative control siRNA for 6 h. L‐M, The semi‐quantification of the proteins in panel H, respectively. All experiments were repeated at least three times, and data are expressed as mean ± SEM, * P < 0.05 vs Ctrl group, # P < 0.05 vs LPS group

Article Snippet: Control siRNA is a non‐targeting 20‐25 nt siRNA (Scramble; Obio technology; China) designed as a negative control.

Techniques: Knockdown, Staining, Transfection, Negative Control, Western Blot, Expressing

Journal: Journal of Cellular and Molecular Medicine

Article Title: YAP accelerates vascular senescence via blocking autophagic flux and activating mTOR

doi: 10.1111/jcmm.15902

Figure Lengend Snippet: Impaired autophagy occurred during YAP promoting cellular and vascular senescence. A, Western blot was preformed to analyse the expression of Beclin1, LC3Ⅱ, LC3Ⅰ and P62 in HUVECs treated with or without 0.1 mg/mL verteporfin in the presence of 0.5 μg/mL LPS for 6 d. B, The semi‐quantifications of the proteins of plane A, respectively. C, Western blot was used to analyse YAP, p‐mTOR (Ser2448), mTOR, Beclin1, LC3Ⅱ, LC3Ⅰ and P62 expression in HUVECs with or without YAP siRNA (siYAP#1, siYAP#2, siYAP#3 for 6 h) in the presence of LPS (0.5 μg/mL LPS for 6 d). D, The semi‐quantifications of the proteins of plane C, respectively. E, Western blot was preformed to analyse p‐mTOR (Ser2448), mTOR, LC3Ⅱ, LC3Ⅰ and P62 in isolated vascular tissue with or without YAP siRNA (siYAP#1, siYAP#2, siYAP#3 for 6 h) in the presence of LPS (0.5 μg/mL for 6 d). F, The semi‐quantifications of the proteins of plane E, respectively. All experiments were repeated at least three times, and data are expressed as mean ± SEM, * P < 0.05 vs Ctrl group, # P < 0.05 vs LPS group

Article Snippet: Control siRNA is a non‐targeting 20‐25 nt siRNA (Scramble; Obio technology; China) designed as a negative control.

Techniques: Western Blot, Expressing, Isolation

Journal: Journal of Cellular and Molecular Medicine

Article Title: YAP accelerates vascular senescence via blocking autophagic flux and activating mTOR

doi: 10.1111/jcmm.15902

Figure Lengend Snippet: Autophagic flux was impaired during YAP promoting cellular senescence. A, Western blot was preformed to analyse the expression of LC3Ⅱ, LC3Ⅰ, P62, YAP, P53, P21 and P16 in HUVECs co‐treated with or without Chloroquine (CQ, 10 mol/L for 24 h) in the presence of YAP siRNA (siYAP#1, siYAP#2, siYAP#3 for 6 h). Experiments were repeated three times. B, The semi‐quantifications of the proteins of plane A, respectively. Data are expressed as mean ± SEM, * P < 0.05 vs Ctrl group, # P < 0.05 vs LPS group. C, Western blot was preformed to analyse the expression of LC3Ⅱ, LC3Ⅰ, P62, YAP, P53 and P16 in HUVECs co‐treated with or without CQ (10 mol/L for 24 h) in the presence of Ad‐YAP. All experiments were repeated at least three times. D, The semi‐quantifications of the proteins of plane C, respectively. Data are expressed as mean ± SEM, * P < 0.05 vs Ctrl group, # P < 0.05 vs Ad‐YAP group. E, Tandem fluorescent mRFP‐GFP‐LC3 adenovirus was subjected to HUVECs to detect the numbers of APs in the presence and absence of CQ (10 mol/L) for 24 h. The nuclei were labelled with DAPI (blue staining); GFP dots are green; mRFP dots are red; YAP is white (Alexa Fluor 647). Scale bar = 10 μm. Experiments were repeated three times. F, Quantitative analysis of APs (yellow dots) and ALs (red dot) in plan E by counting manually; N = 30‐50 nuclei per group. Data are expressed as mean ± SEM,* P < 0.05 vs Ctrl group, # P < 0.05 vs LPS group

Article Snippet: Control siRNA is a non‐targeting 20‐25 nt siRNA (Scramble; Obio technology; China) designed as a negative control.

Techniques: Western Blot, Expressing, Staining

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