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: Cell Death & Disease

Article Title: Recombinant human arginase induced caspase-dependent apoptosis and autophagy in non-Hodgkin's lymphoma cells

doi: 10.1038/cddis.2013.359

Figure Lengend Snippet: rhArg-induced apoptosis was caspase-dependent in NHL cells. ( a ) Raji and Daudi cells were incubated with 1 IU/ml of rhArg for 24, 48, and 72 h and cell lysates were subjected to Western blotting using anti-caspase-3 and tublin antibodies. ( b–d ) Raji and Daudi cells were incubated with 1 IU/ml of rhArg or in combination with 20 μ M z-VAD-fmk for 48 h. ( b ) The LC3 conversion, PARP, and caspase-3 protein levels were detected by western blot analysis. ( c ) Cell viability was measured by MTT assay. ( d ) Cells were stained with Annexin V/PI, and analyzed by flow cytometry. The percentage of Raji and Daudi cells positive for Annexin V staining were presented in bar charts. ( e and f ) Raji and Daudi cells transfected siRNA against caspase-3 followed by treatment of 1 IU/ml of rhArg for 48 h. ( e ) The low expression of caspase-3 of Raji and Daudi cells resulted in siRNA was detected by western blot. ( f ) The percentage of Raji and Daudi cells positive for Annexin V staining were presented in bar charts. The percentages of positive cells were calculated from triplicates of one representative experiment. Error bars showed standard deviation from triplicates of one typical experiment. Similar results were obtained from three independent experiments. * P <0.05, ** P <0.01

Article Snippet: siRNA against Atg5 (sense sequence: 5′-GUGAGAUAUGGUUUGAAUA-3′; antisense sequence: 5′-CACUCUAUACCAAACUUAU-3′), against Beclin-1 (sense sequence: 5′-CAGUUUGGCACAAUCAAUA-3′; antisense sequence: 5′-GUCAAACCGUGUUAGUUAU-3′), against caspase-3 (sense sequence: 5′-AGUGAAGCAAAUCAGAAAC-3′; antisense sequence: 5′-UCACUUCGUUUAGUCUUUG-3′), against ASS (sense sequence: 5′-GCUAUGACGUCAUUGCCUA-3′; antisense sequence: 5′-CGAUACUGCAGUAACGGAU-3′), and a negative control siRNA were purchasedd from Guangzhou RiboBio Co., Ltd (Guangzhou, China).

Techniques: Incubation, Western Blot, MTT Assay, Staining, Flow Cytometry, Transfection, Expressing, Standard Deviation

Journal: Cell Death & Disease

Article Title: Recombinant human arginase induced caspase-dependent apoptosis and autophagy in non-Hodgkin's lymphoma cells

doi: 10.1038/cddis.2013.359

Figure Lengend Snippet: Autophagy played a critical role in rhArg-induced NHL cell death. ( a and b ) Raji and Daudi cells were treated with 1 IU/ml of rhArg, either alone or in combination with 2 mM 3-MA or 20 μ M CQ, for 24 h. ( a ) The cell viability was measured by MTT assay and expressed with optical density (OD)values. ( b ) Cell lysates were analyzed by western blot analysis. ( c ) Lymphoma cells were treated with 1 IU/ml of rhArg, either alone or in combination with 2 mM 3-MA or 20 μ M CQ, for 48 h (Raji) or 24 h (Daudi). Raji and Daudi cells were stained for Annexin V/PI and analyzed by flow cytometry. The percentage of Annexin V-positive cells was presented in bar charts. The percentages of positive cells were calculated from triplicates of one representative experiment. ( d and e ) Raji and Daudi cells were transiently transfected with Atg5 or Beclin-1 siRNA followed by treatment of 1 IU/ml of rhArg for 48 h (Raji) or 24 h (Daudi). Raji ( d ) and Daudi ( e ) cell lysates were subjected to western blot analysis to assess the protein level of Atg5, Beclin-1, and PARP and cell viability was measured by MTT assay. Error bars showed standard deviation from triplicates of one typical experiment. Similar results were obtained from three independent experiments. * P <0.05, ** P <0.01

Article Snippet: siRNA against Atg5 (sense sequence: 5′-GUGAGAUAUGGUUUGAAUA-3′; antisense sequence: 5′-CACUCUAUACCAAACUUAU-3′), against Beclin-1 (sense sequence: 5′-CAGUUUGGCACAAUCAAUA-3′; antisense sequence: 5′-GUCAAACCGUGUUAGUUAU-3′), against caspase-3 (sense sequence: 5′-AGUGAAGCAAAUCAGAAAC-3′; antisense sequence: 5′-UCACUUCGUUUAGUCUUUG-3′), against ASS (sense sequence: 5′-GCUAUGACGUCAUUGCCUA-3′; antisense sequence: 5′-CGAUACUGCAGUAACGGAU-3′), and a negative control siRNA were purchasedd from Guangzhou RiboBio Co., Ltd (Guangzhou, China).

Techniques: MTT Assay, Western Blot, Staining, Flow Cytometry, Transfection, Standard Deviation

Journal: Oncotarget

Article Title: Microfluidic hydrodynamic focusing synthesis of polymer-lipid nanoparticles for siRNA delivery

doi: 10.18632/oncotarget.18281

Figure Lengend Snippet: Properties of nanoparticles prepared by MF and BM methods

Article Snippet: VEGF siRNA (sense sequence: 5′-GGAGUACCCUGAUGAGAUCdTdT-3′; antisense sequence: 5′-GAUCUCAUCAGGGUACUCCdTdT-3′) and 5′-FAM-siRNA were provided by Guangzhou RiboBio Co., Ltd. (Guangzhou, China).

Techniques: Zeta Potential Analyzer

Journal: Oncotarget

Article Title: Microfluidic hydrodynamic focusing synthesis of polymer-lipid nanoparticles for siRNA delivery

doi: 10.18632/oncotarget.18281

Figure Lengend Snippet: Cells were incubated with ( A ) blank P/LNPs or ( B ) siRNA-loaded P/LNPS for 12h, 24 h, 48 h or 72 h in an atmosphere containing 5% CO 2 at37°C. All values were presented as mean ± SD ( * indicates p < 0.05, ** indicates p < 0.01 and *** indicates p < 0.001). Abbreviations: blank P/LNPs-BM, P/LNPs-BM without siRNA, blank P/LNPs-MF, P/LNPs-MF without siRNA, P/LNPs-siRNA-BM, siRNA-loaded P/LNPs-BM, P/LNPs-siRNA-MF, siRNA-loaded P/LNPs-MF.

Article Snippet: VEGF siRNA (sense sequence: 5′-GGAGUACCCUGAUGAGAUCdTdT-3′; antisense sequence: 5′-GAUCUCAUCAGGGUACUCCdTdT-3′) and 5′-FAM-siRNA were provided by Guangzhou RiboBio Co., Ltd. (Guangzhou, China).

Techniques: Incubation

Journal: Oncotarget

Article Title: Microfluidic hydrodynamic focusing synthesis of polymer-lipid nanoparticles for siRNA delivery

doi: 10.18632/oncotarget.18281

Figure Lengend Snippet: The analysis of P/LNPs mediated siRNA delivery in HepG-2 cells was performed by confocal microscopy. Cell nuclei were counterstained with DAPI. Abbreviations: P/LNPs-siRNA-MF, Cy3-siRNA-loaded P/LNPs-MF, P/LNPs-siRNA-BM, Cy3-siRNA-loaded P/LNPs-BM, Cy3-siRNA, Cy3-labeled siRNA.

Article Snippet: VEGF siRNA (sense sequence: 5′-GGAGUACCCUGAUGAGAUCdTdT-3′; antisense sequence: 5′-GAUCUCAUCAGGGUACUCCdTdT-3′) and 5′-FAM-siRNA were provided by Guangzhou RiboBio Co., Ltd. (Guangzhou, China).

Techniques: Confocal Microscopy, Labeling

Journal: Oncotarget

Article Title: Microfluidic hydrodynamic focusing synthesis of polymer-lipid nanoparticles for siRNA delivery

doi: 10.18632/oncotarget.18281

Figure Lengend Snippet: ( A ) Standard curve of Fluorescence Intensity versus Concentration of FAM-siRNA. ( B ) Plasma concentrations of P/LNPs-siRNA-MF and free FAM-siRNA after i.v. administration. The data are presented as means ± SD ( n = 3). Abbreviations: P/LNPs-siRNA-MF, FAM-siRNA-loaded P/LNPs-MF; FAM-siRNA, FAM-labeled siRNA.

Article Snippet: VEGF siRNA (sense sequence: 5′-GGAGUACCCUGAUGAGAUCdTdT-3′; antisense sequence: 5′-GAUCUCAUCAGGGUACUCCdTdT-3′) and 5′-FAM-siRNA were provided by Guangzhou RiboBio Co., Ltd. (Guangzhou, China).

Techniques: Fluorescence, Concentration Assay, Clinical Proteomics, Labeling

Journal: Oncotarget

Article Title: Microfluidic hydrodynamic focusing synthesis of polymer-lipid nanoparticles for siRNA delivery

doi: 10.18632/oncotarget.18281

Figure Lengend Snippet: Plasma pharmacokinetic parameters

Article Snippet: VEGF siRNA (sense sequence: 5′-GGAGUACCCUGAUGAGAUCdTdT-3′; antisense sequence: 5′-GAUCUCAUCAGGGUACUCCdTdT-3′) and 5′-FAM-siRNA were provided by Guangzhou RiboBio Co., Ltd. (Guangzhou, China).

Techniques: Clinical Proteomics

Journal: Oncotarget

Article Title: Microfluidic hydrodynamic focusing synthesis of polymer-lipid nanoparticles for siRNA delivery

doi: 10.18632/oncotarget.18281

Figure Lengend Snippet: ( A ) Tumor volumes were measured by caliper three times a week after P/LNPs-siRNA-BM, P/LNPs-siRNA-MF, or saline treatment. ( B ) Images of the harvested tumors at the time of sacrifice. ( C ) Body weights of mice were monitored during the treatment. ( D ) The average weight of tumors collected at the end of treatment. ( * p < 0.05, ** p < 0.01, *** p < 0.001). Abbreviations: P/LNPs-siRNA-BM, siRNA-loaded P/LNPs-BM; P/LNPs-siRNA-MF, siRNA-loaded P/LNPs-MF.

Article Snippet: VEGF siRNA (sense sequence: 5′-GGAGUACCCUGAUGAGAUCdTdT-3′; antisense sequence: 5′-GAUCUCAUCAGGGUACUCCdTdT-3′) and 5′-FAM-siRNA were provided by Guangzhou RiboBio Co., Ltd. (Guangzhou, China).

Techniques: Saline

Journal: Oncotarget

Article Title: Microfluidic hydrodynamic focusing synthesis of polymer-lipid nanoparticles for siRNA delivery

doi: 10.18632/oncotarget.18281

Figure Lengend Snippet: Notes: ( A ) The level of VEGF mRNA determined by qRT-PCR. ( B ) VEGF protein expression determined via Western blot analysis. Data represents mean ± SD ( n = 3). * indicates p < 0.05, ** indicates p < 0.01 and. *** indicates p < 0.001. Abbreviations: P/LNPs-siRNA-BM, siRNA-loaded P/LNPs-BM; P/LNPs-siRNA-MF, siRNA-loaded P/LNPs-MF.

Article Snippet: VEGF siRNA (sense sequence: 5′-GGAGUACCCUGAUGAGAUCdTdT-3′; antisense sequence: 5′-GAUCUCAUCAGGGUACUCCdTdT-3′) and 5′-FAM-siRNA were provided by Guangzhou RiboBio Co., Ltd. (Guangzhou, China).

Techniques: Quantitative RT-PCR, Expressing, Western Blot

Journal: Oncotarget

Article Title: Microfluidic hydrodynamic focusing synthesis of polymer-lipid nanoparticles for siRNA delivery

doi: 10.18632/oncotarget.18281

Figure Lengend Snippet: Heart, Liver, Spleen, Lung, Kidney, Tumor tissue were collected and stained by hematoxylin/eosin (H&E) after the last treatment. Abbreviations: P/LNPs-siRNA-BM, siRNA-loaded P/LNPs-BM; P/LNPs-siRNA-MF, siRNA-loaded P/LNPs-MF.

Article Snippet: VEGF siRNA (sense sequence: 5′-GGAGUACCCUGAUGAGAUCdTdT-3′; antisense sequence: 5′-GAUCUCAUCAGGGUACUCCdTdT-3′) and 5′-FAM-siRNA were provided by Guangzhou RiboBio Co., Ltd. (Guangzhou, China).

Techniques: Staining

Journal: Oncotarget

Article Title: Microfluidic hydrodynamic focusing synthesis of polymer-lipid nanoparticles for siRNA delivery

doi: 10.18632/oncotarget.18281

Figure Lengend Snippet: In vivo toxicity assay by measuring the levels of ( A ) AST, ( B ) ALT, ( C ) BUN and ( D ) creatinine in serum of mice 48 h after intravenous injections of P/LNPs. Normal ranges of parameters are presented. Data are expressed as mean ± SD ( n = 3). Abbreviation: AST, aspartate aminotransferase; ALT: alanine aminotransferase; BUN: blood urea nitrogen; P/LNPs-siRNA-BM, siRNA-loaded P/LNPs-BM; P/LNPs-siRNA-MF, siRNA-loaded P/LNPs-MF.

Article Snippet: VEGF siRNA (sense sequence: 5′-GGAGUACCCUGAUGAGAUCdTdT-3′; antisense sequence: 5′-GAUCUCAUCAGGGUACUCCdTdT-3′) and 5′-FAM-siRNA were provided by Guangzhou RiboBio Co., Ltd. (Guangzhou, China).

Techniques: In Vivo

Journal: Oncotarget

Article Title: Microfluidic hydrodynamic focusing synthesis of polymer-lipid nanoparticles for siRNA delivery

doi: 10.18632/oncotarget.18281

Figure Lengend Snippet: The device consisted of three inlet ports and one outlet port and was control by two syringe pumps. siRNA solutions were introduced from inlets 1 and 3, and the lipid solution was injected from inlet 2 and was mixed with siRNA solution through an S-style MF channel at 0.8 mL/min flow rate. Resulting P/LNPs was collected at the outlet port, followed by sonication and dialysis.

Article Snippet: VEGF siRNA (sense sequence: 5′-GGAGUACCCUGAUGAGAUCdTdT-3′; antisense sequence: 5′-GAUCUCAUCAGGGUACUCCdTdT-3′) and 5′-FAM-siRNA were provided by Guangzhou RiboBio Co., Ltd. (Guangzhou, China).

Techniques: Control, Injection, Sonication

Journal: Molecular Pain

Article Title: Upregulation of bone morphogenetic protein 2 ( Bmp2 ) in dorsal root ganglion in a rat model of bone cancer pain

doi: 10.1177/1744806918824250

Figure Lengend Snippet: Targeting Bmp2 by intrathecal siRNA administration in BCP rats. (a) The knockdown effect of Bmp2 siRNA was tested by RT-qPCR ( n =8). (b) and (c) PWMT and PWTL were measured after Bmp2 siRNA administration in BCP rats at different time points. Data (b and c) are presented as mean ± SD ( n =12 rats per group), and inter-group differences are assessed using Student’s t test. * P <0.05, ** P <0.01, compared with the BCP+NC-siRNA group. BCP: bone cancer pain; BMP2: bone morphogenetic protein 2.

Article Snippet: Bmp2 was targeted by siRNA (sequence: Sense: 5′-GAAGCCAUCGAGGAACUUUTT-3′, Antisense: 5′-AAAGUUCCUCGAUGGCUUCTT-3′; GenePharma, Shanghai, China).

Techniques: Knockdown, Quantitative RT-PCR

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