Journal: eLife

Article Title: Cerebrospinal fluid-contacting neuron tracing reveals structural and functional connectivity for locomotion in the mouse spinal cord

doi: 10.7554/elife.83108

Figure Lengend Snippet: Figure 4. Serial block-face scanning electron microscopy (SBF-SEM) analyses with COX4-dAPEX2 and SYP-HRP labeling reveal structures and recurrent connections of cerebrospinal fluid-contacting neurons (CSF-cNs). (A, B) Representative light microscopic images of 3, 3'-diaminobenzidine (DAB)- stained spinal cord sections of Pkd2l1-Cre mice with injection of adeno-associated virus (AAV)-Syn-DIO-COX4-dAPEX2 (A) or AAV-Syn-DIO-SYP-HRP (B). (C) A low-magnification electron microscopic image of the cervical cord around the central canal (Cc) in Pkd2l1-Cre mice injected with AAV-Syn-DIO-

Article Snippet: AAV plasmids were generated as follows: for pAAV- Syn- mCherry, mCherry of pmCherry- C1 plasmid (Clontech) was subcloned into pAAV- hSyn- EGFP (Addgene, #50465) in a substitution of EGFP; for pAAV- Syn- DIO- COX4- dAPEX2 and pAAV- Syn- DIO- SYP- HRP, COX4- dAPEX2 and SYP- HRP derived from pAAV- EF1a- COX4- dAPEX2 (Addgene, #117176) and pAAV- EF1a- SYP- HRP (Addgene, #117185) were subcloned into pAAV- Syn- DIO- MCS that had multicloning sites (MCS) between the DIO site of pAAV- Syn- DIO- mCherry (Addgene, #50459) in a substitution of mCherry; and for pAAV- Syn- DIOChR2- mCherry, ChR2(H134R)- mCherry derived from pAAV- CAG- ChR2(H134R)- mCherry (Addgene, #100054) was subcloned into pAAV- Syn- DIO- MCS plasmid. pAAV- hSyn- EGFP (Addgene, #50465), pAAV- CAG- EGFP (pENN.AAV.CB7.CI.eGFP.WPRE.rBG; Penn vector core, #AV- 1- PV1963), and pAAVSyn- DIO- mCherry (Addgene, #50459) were further used for AAV production.

Techniques: Blocking Assay, Electron Microscopy, Labeling, Staining, Injection, Virus

Journal: Science translational medicine

Article Title: Insights into neuroepigenetics through human histone deacetylase PET imaging

doi: 10.1126/scitranslmed.aaf7551

Figure Lengend Snippet: Human neural progenitor cells were treated with DMSO, Martinostat (MSTAT 0.5 μM, 2.5 μM) and SAHA for 24 hrs. (A) Whole cell lysates were prepared (n=3 replicates). Equivalent amounts of total protein were compared through western blotting (Fig. S15A). Histone acetylation immunoreactive band intensity values were normalized to GAPDH intensity values. Error bars represent standard deviation. Histone H3 lysine 9 and histone H4 lysine 12 acetylation levels were significantly increased as compared to DMSO with 2.5 μM Martinostat and SAHA treatments (repeated measures two-way ANOVA with Dunnett’s multiple comparisons correction, α=0.05, ** p<0.01, **** p<0.0001). (B) RNA was extracted (n=3 replicates) and converted into cDNA. mRNA transcript levels of memory/neuronal plasticity (blue) and monogenic neurological disorder-related genes (green) were compared through qPCR (n=9 technical replicates) and normalized to GAPDH mRNA levels (Fig. S15B). Error bars represent standard error of the mean. BDNF, SYT1, SYP, and GRN levels were significantly increased as compared to DMSO with 2.5 μM Martinostat treatment (repeated measures two-way ANOVA with Dunnett’s multiple comparisons correction, α=0.05, * p<0.05, ** p<0.01, **** p<0.0001). BDNF, SYP, and GRN levels were significantly increased as compared to DMSO with SAHA treatment (repeated measures two-way ANOVA with Dunnett’s multiple comparisons correction, α=0.05, ** p<0.01, **** p<0.0001). Gene abbreviations used are as follows: BDNF, brain-derived neurotrophic factor; CDK5, cyclin-dependent kinase 5; SYT1, synaptotagmin-1; SYP, synaptophysin; GRN, progranulin; FXN, frataxin; GAPDH, glyceraldehyde-3-phosphate dehydrogenase.

Article Snippet: Into each well 5 μL of TaqMan Gene Expression Master Mix (Thermo Scientific #4369510), 0.5 μL of 20X commercial TaqMan primer probe for each gene described (Thermo Scientific: GRN , Hs00963707_g1; FXN , Hs00175940_m1; BDNF , Hs00380947_m1; EGR1 , Hs00152928_m1; CDK5, Hs00358991_g1; SYT1 , Hs00194572_m1; SYP, Hs00300531_m1), 0.5 μL of DNase/RNase free water, and 4 μL of above diluted cDNA was added.

Techniques: Western Blot, Standard Deviation, Derivative Assay

Journal: Drug Design, Development and Therapy

Article Title: Integrating Network Pharmacology and Component Analysis to Study the Potential Mechanisms of Qi-Fu-Yin Decoction in Treating Alzheimer’s Disease

doi: 10.2147/DDDT.S402624

Figure Lengend Snippet: Primer Sequence of Target Gene and Internal Reference Gene

Article Snippet: D-galactose was purchased from Shanghai Macklin Biochemical Co., Ltd. Piracetam was purchased from Tianjin Jinshi Pharmaceutical Co., Ltd; The primary antibodies for Bcl-2, Caspase-1, Bax, PSD95, SYP and β-actin were purchased from Wuhan Boster Biological Technology Co., Ltd. IL-1β, IL-18 and TNF-α ELISA kits were obtained from Elabscience Biotechnology Co., Ltd. DAPI purchased from Beijing Solarbio Science & Technology Co., Ltd.

Techniques: Sequencing

Journal: Drug Design, Development and Therapy

Article Title: Integrating Network Pharmacology and Component Analysis to Study the Potential Mechanisms of Qi-Fu-Yin Decoction in Treating Alzheimer’s Disease

doi: 10.2147/DDDT.S402624

Figure Lengend Snippet: The effect of QFY on synapses and neuronal apoptosis. ( A ) The expression of PSD95 and SYP protein in mice hippocampus. ( B ) Amplification curve, fusion curve and mRNA expression of PSD95 and SYP. ( C ) The expression of Bax, Bcl-2, and Caspase-1 protein in mice hippocampus. n = 5 in each group, ** P < 0.01 vs the con group; ## P < 0.01 vs the model group.

Article Snippet: D-galactose was purchased from Shanghai Macklin Biochemical Co., Ltd. Piracetam was purchased from Tianjin Jinshi Pharmaceutical Co., Ltd; The primary antibodies for Bcl-2, Caspase-1, Bax, PSD95, SYP and β-actin were purchased from Wuhan Boster Biological Technology Co., Ltd. IL-1β, IL-18 and TNF-α ELISA kits were obtained from Elabscience Biotechnology Co., Ltd. DAPI purchased from Beijing Solarbio Science & Technology Co., Ltd.

Techniques: Expressing, Amplification