mef2d sirna  (Santa Cruz Biotechnology)

Journal: Neurobiology of stress

Article Title: LGI1 governs neuritin-mediated resilience to chronic stress.

doi: 10.1016/j.ynstr.2021.100373

Figure Lengend Snippet: Fig. 2. Neuritin induces LGI1 expression through HDAC5 phosphorylation and MEF2D-mediated transcription. (A) Representative immunoblots of p-HDAC5 in hippocampal neurons (DIV7) treated with various concentrations of recombinant soluble neuritin for 30 min or with KCl (30 mM) for 6 h (n = 4–5). (B) Repre sentative immunoblots of p-HDAC5 from hippocampal neurons pretreated with KN-62 (30 μM) or G¨o6976 (1 μM) for 30 min and incubated with soluble neuritin or KCl (n = 3). (C–E) Luciferase assays. MEF2-luiciferase activity was normalized with Renilla luciferase activity and is depicted relative to the control (CTL), and expressed as fold change relative to the CTL. (C) Mouse hippocampal neurons (DIV4) transfected with pGL3-Luc and pGL3-MEF2-Luc were treated with neuritin for the indicated times (n = 4). (D) Mouse hippocampal neurons transfected with pGL3-Luc and PGL3-MEF2-Luc were treated with neuritin in the presence of KN-62 or G¨o6976 (n = 4). (E) Neurons transfected with pGL3-Luc, pGL3-MEF2-Luc, pCl-neo, pCl-neo-HDAC5-WT or pCl-neo-HDAC5-S/A were treated with recombinant soluble neuritin for 1 h (n = 7). (F) ChIP assays. Binding of HDAC5 to the Lgi1 promoter was decreased in recombinant soluble neuritin-treated mouse hippocampal neurons (n = 3). (G) IP. Neurons (DIV4) transfected with Myc-HDAC5 were treated with recombinant soluble neuritin (200 ng/ml) for 1 h. Binding of HDAC5 to MEF2D in mouse hippocampal neurons was decreased by recombinant soluble neuritin treatment (n = 3). (H) Neurons (DIV4) transfected with control siRNA and Mef2d siRNA were treated with recombinant soluble neuritin (200 ng/ml) for 6 h. Representative immunoblots (Left) and quantitative data (Right) for MEF2D or LGI1 protein expression (n = 3). In (A)–(H), Data are represented as mean ± SEM; *p < 0.05, **p < 0.01, ***p < 0.001 compared with CTL, #p < 0.05, ###p < 0.001 compared with recombinant soluble neuritin treatment. Statistics: Student’s t-test (A, C, F and G). One-way ANOVA (B) or two-way ANOVA (D) followed by LSD posttest. Two-way ANOVA followed by Newman-Keuls posttest (E) or Bonferroni posttest (H). Statistics detailed in Table S4.

Article Snippet: Control siRNA (SC-37007, Santa Cruz, TX, USA) and Mef2d siRNA (SC-38065, Santa Cruz) were solubilized in RNAse-free water.

Techniques: Expressing, Phospho-proteomics, Western Blot, Recombinant, Incubation, Luciferase, Activity Assay, Control, Transfection, Binding Assay

Journal: International Journal of Nanomedicine

Article Title: Silencing of MEF2D by siRNA Loaded Selenium Nanoparticles for Ovarian Cancer Therapy

doi: 10.2147/IJN.S270441

Figure Lengend Snippet: The Primer Sequences of MEF2D and GAPDH for Quantitative Real-Time PCR

Article Snippet: The released MEF2D-siRNA was detected for 15 hours by a spectromax quickdrop.

Techniques:

Journal: International Journal of Nanomedicine

Article Title: Silencing of MEF2D by siRNA Loaded Selenium Nanoparticles for Ovarian Cancer Therapy

doi: 10.2147/IJN.S270441

Figure Lengend Snippet: Schematic illustration of the formation of RGDfC-Se@MEF2D-siRNA.

Article Snippet: The released MEF2D-siRNA was detected for 15 hours by a spectromax quickdrop.

Techniques:

Journal: International Journal of Nanomedicine

Article Title: Silencing of MEF2D by siRNA Loaded Selenium Nanoparticles for Ovarian Cancer Therapy

doi: 10.2147/IJN.S270441

Figure Lengend Snippet: ( A ) Zeta potentials examination of SeNPs, RGDfC, R-SeNPs, and R-Se@MEF2D-siRNA. ( B ) MEF2D-siRNA stability in R-Se@MEF2D-siRNA at various N/P ratios was assessed by agarose gel electrophoresis assay.

Article Snippet: The released MEF2D-siRNA was detected for 15 hours by a spectromax quickdrop.

Techniques: Agarose Gel Electrophoresis

Journal: International Journal of Nanomedicine

Article Title: Silencing of MEF2D by siRNA Loaded Selenium Nanoparticles for Ovarian Cancer Therapy

doi: 10.2147/IJN.S270441

Figure Lengend Snippet: The uptake of R-Se@MEF2D-siRNA was visualized by fluorescence microscope. ( A ) R-Se@MEF2D-siRNA was taken up by SKOV3 cells. ( B ) R-Se@MEF2D-siRNA was taken up by HUVEC. Scale bar indicates 20 μm.

Article Snippet: The released MEF2D-siRNA was detected for 15 hours by a spectromax quickdrop.

Techniques: Fluorescence, Microscopy

Journal: International Journal of Nanomedicine

Article Title: Silencing of MEF2D by siRNA Loaded Selenium Nanoparticles for Ovarian Cancer Therapy

doi: 10.2147/IJN.S270441

Figure Lengend Snippet: ( A ) The uptake of R-Se@MEF2D-siRNA in SKOV3 cells was influenced by low temperature and endocytosis inhibitors. * P <0.05, ** P <0.01 vs control. ( B ) The release of MEF2D-siRNA from R-SeNPs at pH5.4 and pH7.4. * P <0.05 vs pH5.4 group. ( C ) Relative expressions of MEF2D in SKOV3 cells were examined using qRT-PCR. ** P <0.01 vs control group.

Article Snippet: The released MEF2D-siRNA was detected for 15 hours by a spectromax quickdrop.

Techniques: Quantitative RT-PCR

Journal: International Journal of Nanomedicine

Article Title: Silencing of MEF2D by siRNA Loaded Selenium Nanoparticles for Ovarian Cancer Therapy

doi: 10.2147/IJN.S270441

Figure Lengend Snippet: ( A ) Cytotoxicity of SKOV3 cells exposed to various concentrations of R-Se@siNC and R-Se@MEF2D-siRNA after 48 hours. ( B ) Cytotoxicity of HUVEC exposed to various concentrations of R-Se@MEF2D-siRNA. ** P <0.01 vs R-Se@siNC. ( C ) Cell cycle phase distribution of nuclear DNA in R-Se@siNC or R-Se@MEF2D-siRNA-treated SKOV3 cells was determined by flow cytometry.

Article Snippet: The released MEF2D-siRNA was detected for 15 hours by a spectromax quickdrop.

Techniques: Flow Cytometry

Journal: International Journal of Nanomedicine

Article Title: Silencing of MEF2D by siRNA Loaded Selenium Nanoparticles for Ovarian Cancer Therapy

doi: 10.2147/IJN.S270441

Figure Lengend Snippet: ( A ) R-Se@MEF2D-siRNA induced changes of MMP in SKOV3 cells. The MMP values of treated cells were examined by flow cytometry using JC-1 staining. ( B ) R-Se@MEF2D-siRNA increased the ROS level of SKOV3 cells. The treated SKOV3 cells were co-incubated with DCFDA for 30 minutes and then visualized by a fluorescence microscope to measure intracellular ROS levels. Scale bar indicates 20 μm.

Article Snippet: The released MEF2D-siRNA was detected for 15 hours by a spectromax quickdrop.

Techniques: Flow Cytometry, Staining, Incubation, Fluorescence, Microscopy

Journal: International Journal of Nanomedicine

Article Title: Silencing of MEF2D by siRNA Loaded Selenium Nanoparticles for Ovarian Cancer Therapy

doi: 10.2147/IJN.S270441

Figure Lengend Snippet: H&E stained heart, kidneys, liver, lung, and spleen tissues from mice treated with saline, R-Se@MEF2D-siRNA or R-Se@siNC. Bar indicates 50 µm.

Article Snippet: The released MEF2D-siRNA was detected for 15 hours by a spectromax quickdrop.

Techniques: Staining