Journal: Journal of Translational Medicine

Article Title: FNDC5 attenuates obesity-induced cardiac hypertrophy by inactivating JAK2/STAT3-associated inflammation and oxidative stress

doi: 10.1186/s12967-019-1857-8

Figure Lengend Snippet: FNDC5 deficiency aggravated HFD-induced inflammation in heart. a Cardiac Tnf - α, Il1b and Il6 mRNA levels determined with qPCR. b Phosphorylation levels of p38 and ERK in heart determined with Western blot. c NFκB activation in heart. N-p65: p65 in nucleus; C-p65: p65 in cytoplasm. Values are mean ± SEM. *P < 0.05 vs. WT. † P < 0.05 vs. Ctrl. n = 3

Article Snippet: The FNDC5 specific siRNA (sense sequence: 5′GAUGGCCUCUAAGAACAAA3′; antisense sequence: 3′CUACCGGAGAUUCUUGUUU5′) was purchased from RiboBio (Guangzhou, China).

Techniques: Western Blot, Activation Assay

Journal: Journal of Translational Medicine

Article Title: FNDC5 attenuates obesity-induced cardiac hypertrophy by inactivating JAK2/STAT3-associated inflammation and oxidative stress

doi: 10.1186/s12967-019-1857-8

Figure Lengend Snippet: FNDC5 deficiency enhanced HFD-induced oxidative stress and phosphorylated JAK2 and STAT3 level in heart. a Superoxide dismutase (SOD) activity and malondialdehyde (MDA) level in heart. b Expression of NAPDH oxidases (NOX2 and NOX4) protein in heart. c Phosphorylated JAK2 and STAT3 level in heart. Values are mean ± SEM. *P < 0.05 vs. WT. † P < 0.05 vs. Ctrl. n = 6

Article Snippet: The FNDC5 specific siRNA (sense sequence: 5′GAUGGCCUCUAAGAACAAA3′; antisense sequence: 3′CUACCGGAGAUUCUUGUUU5′) was purchased from RiboBio (Guangzhou, China).

Techniques: Activity Assay, Expressing

Journal: Journal of Translational Medicine

Article Title: FNDC5 attenuates obesity-induced cardiac hypertrophy by inactivating JAK2/STAT3-associated inflammation and oxidative stress

doi: 10.1186/s12967-019-1857-8

Figure Lengend Snippet: FNDC5 deficiency enhanced palmitate-induced inflammation and NOX4 expression in primary cardiomyocytes (CMs). a Effects of palmitate (PA, 400 μM) treatment on Tnf - α, Il1b and Il6 mRNA levels in CMs of WT and FNDC5 −/− mice. b Effects of PA treatment on NOX2 and NOX4 protein level in CMs of WT and FNDC5 −/− mice. The measurement was 24 h after PA treatment for measuring the mRNA levels or protein levels. PA: Palmitate; Veh: Vehicle; F −/− : FNDC5 −/− . Values are mean ± SEM. *P < 0.05 vs. Veh. † P < 0.05 vs. CMs-WT. n = 3

Article Snippet: The FNDC5 specific siRNA (sense sequence: 5′GAUGGCCUCUAAGAACAAA3′; antisense sequence: 3′CUACCGGAGAUUCUUGUUU5′) was purchased from RiboBio (Guangzhou, China).

Techniques: Expressing

Journal: Journal of Translational Medicine

Article Title: FNDC5 attenuates obesity-induced cardiac hypertrophy by inactivating JAK2/STAT3-associated inflammation and oxidative stress

doi: 10.1186/s12967-019-1857-8

Figure Lengend Snippet: Exogenous FNDC5 pretreatment attenuated palmitate-induced inflammation and oxidative stress in CMs. a , b Effects of exogenous FNDC5 pretreatment on cardiac hypertrophy markers ( Nppa, Nppb and Myh7 ) and inflammation markers ( Tnf - α, Il1b and Il6 ) mRNA levels in PA-stimulated CMs. c Effects of exogenous FNDC5 pretreatment on NO production in cell culture supernatant of PA-stimulated CMs. d Effects of exogenous FNDC5 pretreatment on NOX expression in PA-stimulated CMs. PA: Palmitate; Veh: Vehicle. Values are mean ± SEM. *P < 0.05 vs. Veh. † P < 0.05 vs. PA. n = 6

Article Snippet: The FNDC5 specific siRNA (sense sequence: 5′GAUGGCCUCUAAGAACAAA3′; antisense sequence: 3′CUACCGGAGAUUCUUGUUU5′) was purchased from RiboBio (Guangzhou, China).

Techniques: Cell Culture, Expressing

Journal: Journal of Translational Medicine

Article Title: FNDC5 attenuates obesity-induced cardiac hypertrophy by inactivating JAK2/STAT3-associated inflammation and oxidative stress

doi: 10.1186/s12967-019-1857-8

Figure Lengend Snippet: FNDC5 overexpression (OE) attenuated cardiac hypertrophy, inflammation and oxidative stress in HFD-fed mice. a , b Serum and heart/muscle FNDC5 levels after lentiviral vector-mediated FNDC5 overexpression. c Cardiomyocyte area in heart determined by H&E staining. d , e mRNA levels of the cardiac hypertrophy markers ( Nppa, Nppb and Myh7 ) and inflammation markers ( Tnf - α, Il1b and Il6 ). f SOD activity and MDA level in heart. g NOX2, NOX4 level and NFκB inactivation in heart. h Phosphorylation levels of p38 and ERK in heart. Values are mean ± SEM. *P < 0.05 vs. vector. n = 6

Article Snippet: The FNDC5 specific siRNA (sense sequence: 5′GAUGGCCUCUAAGAACAAA3′; antisense sequence: 3′CUACCGGAGAUUCUUGUUU5′) was purchased from RiboBio (Guangzhou, China).

Techniques: Over Expression, Plasmid Preparation, Staining, Activity Assay

Journal: Journal of Translational Medicine

Article Title: FNDC5 attenuates obesity-induced cardiac hypertrophy by inactivating JAK2/STAT3-associated inflammation and oxidative stress

doi: 10.1186/s12967-019-1857-8

Figure Lengend Snippet: Schematic illustrates a possible mechanism of FNDC5 in obesity-induced cardiac hypertrophy. Obesity-induced lipid overload has a great impact on the production of reactive oxygen species (ROS) and upregulation of TNF-α, IL-1β and IL-6 levels, which leads to cardiac inflammation and oxidative stress. The enhanced cardiac inflammation and oxidative stress are involved in pathogenesis of cardiac injury and remodeling. FNDC5 may exert its protective function on the enhanced inflammation and oxidative stress via JAK2/STAT3 pathway through an unknown receptor

Article Snippet: The FNDC5 specific siRNA (sense sequence: 5′GAUGGCCUCUAAGAACAAA3′; antisense sequence: 3′CUACCGGAGAUUCUUGUUU5′) was purchased from RiboBio (Guangzhou, China).

Techniques:

Journal: Journal of Translational Medicine

Article Title: FNDC5 attenuates obesity-induced cardiac hypertrophy by inactivating JAK2/STAT3-associated inflammation and oxidative stress

doi: 10.1186/s12967-019-1857-8

Figure Lengend Snippet: Echocardiographic assessment of left ventricle functions in mice

Article Snippet: The FNDC5 specific siRNA (sense sequence: 5′GAUGGCCUCUAAGAACAAA3′; antisense sequence: 3′CUACCGGAGAUUCUUGUUU5′) was purchased from RiboBio (Guangzhou, China).

Techniques:

Journal: Frontiers in Neuroscience

Article Title: Improving the Delivery of SOD1 Antisense Oligonucleotides to Motor Neurons Using Calcium Phosphate-Lipid Nanoparticles

doi: 10.3389/fnins.2017.00476

Figure Lengend Snippet: Physical properties of SOD1-ASO-loaded CaP-lipid NPs with a Ca/P ratio 100:1. Schematic representation of SOD1-ASO loaded CaP-lipid NP formulation (A) , particle size (d.nm) of SOD1-ASO loaded (black) and non-loaded (red) CaP-lipid NPs determined by DLS (B) , zeta potential of SOD1-ASO CaP-lipid NP (C) , transmission electron micrograph (TEM) of SOD1-ASO CaP-lipid NPs visualized by negative staining (D) , pH-sensitivity as indicated by change in number size (d.nm) of CaP-lipid NPs in PBS at pH 7.4, 6.5, and 5.0 following incubation at 37°C for up to 4 h (E) . ** P < 0.01.

Article Snippet: SOD1 antisense oligonucleotide (target sequence 5′-CCG TCG CCC TTC AGC ACG CA-3′) (Patent US 20120214865 A1) and a scrambled negative control oligonucleotide (target sequence 5'-GCC AGC CTA CGA CTC CGC TC-3′) were synthesized by GeneWorks.

Techniques: Formulation, Zeta Potential Analyzer, Transmission Assay, Negative Staining, Incubation

Journal: Frontiers in Neuroscience

Article Title: Improving the Delivery of SOD1 Antisense Oligonucleotides to Motor Neurons Using Calcium Phosphate-Lipid Nanoparticles

doi: 10.3389/fnins.2017.00476

Figure Lengend Snippet: Particle size, Zeta (ζ)-potential, polydispersity index, and encapsulation effiency of SOD1 antisense oligonucleotide calcium phosphate-lipid nanoparticles (SOD1-ASO CaP-lipid NPs) prepared at different CaCl 2 , and Na 2 HPO 4 concentrations.

Article Snippet: SOD1 antisense oligonucleotide (target sequence 5′-CCG TCG CCC TTC AGC ACG CA-3′) (Patent US 20120214865 A1) and a scrambled negative control oligonucleotide (target sequence 5'-GCC AGC CTA CGA CTC CGC TC-3′) were synthesized by GeneWorks.

Techniques: Encapsulation, Zeta Potential Analyzer, Concentration Assay

Journal: Frontiers in Neuroscience

Article Title: Improving the Delivery of SOD1 Antisense Oligonucleotides to Motor Neurons Using Calcium Phosphate-Lipid Nanoparticles

doi: 10.3389/fnins.2017.00476

Figure Lengend Snippet: Internalization of CaP-lipid NPs into NSC-34 cells and knockdown of SOD1 protein expression in HEK293 cells. The CaP nanoparticles contained LissRdB-DSPE (Ex = 560 nm; Em = 583 nm) in the outer lipid layer. NSC-34 cells were incubated at 37°C with LissRdB-DSPE CaP-lipid NPs diluted either 1:5 or 1:10 (v/v) for 30, 60 (A) and 90 min (B) . Images were acquired with a 63 × objective (512 × 512 pixels; physical length 246.03 μm × 246.03 μm). Change in average cell fluorescence over 90 min incubation period (C) . Red dotted line represents auto fluorescence of NSC-34 cells for comparison. *** P < 0.001 and # P < 0.05 signifies significant difference compared to respective 30 min data. HEK293 cells were treated with SOD1-ASO CaP-lipid NPs for 72 h at 37°C (Lane 3). Control treatments included non-loaded CaP-lipid NPs (Lane 2) and non-loaded CaP-lipid NPs with SOD1-ASO free in solution (Lane 1). Controls using Lipofectamine 2000 Transfection Reagent with SOD1-ASO or scrambled negative SOD1 oligonucleotide are not shown here. The change in SOD1 protein expression (16 kDa) was visualized sing SuperSignal West Pico Chemiluminescent Substrate and exposed using an Amersham 600RGB Imager (D) .

Article Snippet: SOD1 antisense oligonucleotide (target sequence 5′-CCG TCG CCC TTC AGC ACG CA-3′) (Patent US 20120214865 A1) and a scrambled negative control oligonucleotide (target sequence 5'-GCC AGC CTA CGA CTC CGC TC-3′) were synthesized by GeneWorks.

Techniques: Knockdown, Expressing, Incubation, Fluorescence, Comparison, Control, Transfection

Journal: iScience

Article Title: Divergent input patterns to the central lateral amygdala play a duet in fear memory formation

doi: 10.1016/j.isci.2024.110886

Figure Lengend Snippet: Modulation of CB1Rs in the LPB Glu →CeL SOM pathway affects conditioned fear memory acquisition (A) Schematic of the in vitro slice recording paradigm. (B) Representative average traces of eEPSCs in the absence (top) and presence of the CB1R agonist WIN55,212-2 (middle) or the CB1R antagonist AM251 (bottom). (C) Bath application of WIN55,212-2 significantly decreased the amplitude of eEPSCs, whereas AM251 significantly increased the amplitude of eEPSCs compared with the baseline (one-way ANOVA, F (2,12) = 57.97, ∗∗∗∗ p < 0.0001; Tukey’s post hoc test, Baseline versus WIN55, ∗∗∗ p = 0.0004; Baseline versus AM251, ∗∗∗ p = 0.0005; WIN55 versus AM251, ∗∗∗∗ p < 0.0001; n = 5 neurons from 3 mice for each group). (D) Schematic of the injection paradigm to test whether CB1Rs in the CeL control fear memory acquisition. (E) Mice with CeL injections of the CB1R agonist WIN55 exhibited similar fear responses across the duration of fear conditioning as the other two groups (two-way ANOVA, group: F (2,21) = 0.5255, p = 0.5988; time: F (3,63) = 93.78, ∗∗∗∗ p < 0.0001; interaction: F (6,63) = 0.824, p = 0.5556; n = 8 mice). (F and G) Microinjection of the agonist WIN55 into the CeL significantly decreased freezing levels in the contextual (F, one-way ANOVA, F (2,21) = 11.52, ∗∗∗ p = 0.0004; Tukey’s post hoc test, Vehicle versus WIN55, ∗∗∗ p = 0.0004; WIN55 versus WIN55+AM251, ∗ p = 0.0101; n = 8 mice) and cued (G, two-way ANOVA, group: F (2,21) = 2.892, p = 0.0777; time: F (1,21) = 214.3, ∗∗∗∗ p < 0.0001; interaction: F (2,21) = 5.214, ∗ p = 0.0145; Bonferroni’s post hoc test, freezing for Vehicle versus WIN55 during Tone presentation, ∗∗ p = 0.0014; n = 8 mice) fear memory tests, and the effect of WIN55 could be reversed with administration of the CB1R antagonist AM251. (H and I) The expression levels of CB1R mRNA in the LPB (H, two-sided unpaired t -test, t (1/10) = 4.282, ∗∗ p = 0.0016, n = 6 mice) and CB1R protein in the CeL (I, two-sided unpaired t -test, t (1/10) = 4.426, ∗∗ p = 0.0013, n = 6 mice) both decreased significantly after LPB injection of siRNA targeting CB1Rs. (J) Time course and schematic of the injection paradigm to identify the effects of CB1Rs at the LPB→CeL terminals on fear memory acquisition. (K) There were no significant differences among the groups in freezing levels during the fear conditioning session (two-way ANOVA, group: F (3,28) = 0.3493, p = 0.79; time: F (3,84) = 98.5, ∗∗∗∗ p < 0.0001; interaction: F (9,84) = 0.4148, p = 0.9239. n = 8 mice). (L and M) Effects of selective CB1R knockdown in the LPB on CB1R agonist-induced contextual (L, one-way ANOVA, F (3,28) = 8.272, ∗∗∗ p = 0.0004; Bonferroni’s post hoc test, NC + Vehicle versus NC + WIN55, ∗∗ p = 0.0011; NC + WIN55 versus siRNA+Vehicle, ∗∗ p = 0.0012; n = 8 mice) and cued (M, two-way ANOVA, F (3,28) = 12.8, ∗∗∗∗ p < 0.0001; Bonferroni’s post hoc test, before Tone presentation: For NC + Vehicle versus siRNA+Vehicle, ∗ p = 0.033; For NC + WIN55 versus siRNA+Vehicle, ∗∗∗ p = 0.0007; For siRNA +WIN55 versus siRNA+Vehicle, ∗ p = 0.011; during Tone presentation: For NC + Vehicle versus NC + WIN55, ∗∗∗ p = 0.0007; For siRNA+Vehicle versus NC + WIN55, ∗∗∗ p = 0.0003; n = 8 mice) fear memory impairment. All of the data are presented as mean ± SEM. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: SiRNA sequence : CB1R Sense : GACAUUCAGUACGAAGAUATT Antisense : UAUCUUCGUACUGAAUGUCTT , Genepharma , N/A.

Techniques: In Vitro, Injection, Control, Microinjection, Expressing, Knockdown

Journal: iScience

Article Title: Divergent input patterns to the central lateral amygdala play a duet in fear memory formation

doi: 10.1016/j.isci.2024.110886

Figure Lengend Snippet:

Article Snippet: SiRNA sequence : CB1R Sense : GACAUUCAGUACGAAGAUATT Antisense : UAUCUUCGUACUGAAUGUCTT , Genepharma , N/A.

Techniques: Virus, Recombinant, Protein Extraction, Purification, Sequencing, Negative Control, Software