Journal: Journal of Neuroscience

Article Title: REST Regulates Non-Cell-Autonomous Neuronal Differentiation and Maturation of Neural Progenitor Cells via Secretogranin II

doi: 10.1523/jneurosci.4286-14.2015

Figure Lengend Snippet: Figure 4. Scg2 upregulation is required downstream of REST for differentiation of AHPs. A, REST-Scg2 double knockdown rescues REST knockdown phenotype. Immunofluorescent staining of AHPs transfected with shSCR/shSCR, shSCR/shREST, shSCR/ shScg2,orshREST/shScg2culturedinthepresenceofFGF2for4d.GreenrepresentscellsexpressingshRNA.BluerepresentsDAPI. Red represents TUJ1. B, REST/Scg2 double knockdown results in decreased neuronal differentiation compared with REST knock- down alone, as quantified by percentage of GFP /-tubulin III cells. ***p 5.68 10 7.

Article Snippet: Recombinant SCG2 (MW 70.8 kDa, NP_003460) was purchased from Origene and used at 20 ng/ml, resuspended in 0.1% BSA in PBS.

Techniques: Knockdown, Staining, Transfection

Journal: Journal of Neuroscience

Article Title: REST Regulates Non-Cell-Autonomous Neuronal Differentiation and Maturation of Neural Progenitor Cells via Secretogranin II

doi: 10.1523/jneurosci.4286-14.2015

Figure Lengend Snippet: Figure 5. Exogenous SCG2 induces differentiation and maturation of AHPs. A, Recombinant SCG2 was added in the top chamber (treatment chamber) of a microfluidic device; WT AHPs were plated in the bottom chamber (cell chamber). B, Effect of recombinant SCG2 treatment (20 ng/ml) on AHPs. Recombinant SCG2-treated or untreated AHPs were stained with anti-TUJ1 (green), anti-Map2 (red), and DAPI (blue). C, Treatment with SCG2 promoted significant neurite outgrowth. ***p 0.0005. n 80. D, SCG2 promoted neurite branching of wild-type AHPs. The number ofneuritesub-branchesincreasedupontreatmentwithrecombinantSCG2.E,ExampleofmEPSCrecordingstakenfromshSCR_SCG2-andshREST_SCG2-treatedneurons.F,TreatingshSCRprimary neuronswithSCG2(10ng/ml)wassufficienttoincreasemEPSCamplitudetoalevelcomparablewiththatofshREST-transfectedneurons(shSCR_SCG2:n13,mean17.0 2.1pA;shREST_SCG2: n8,mean19.0 1.7pA).G,SCG2treatmentalsoabolishedthedifferenceinmEPSCfrequencybetweenshSCR-andshREST-transfectedneurons(shSCR_SCG2:mean6.4 2.0Hz;shREST_SCG2: mean5.8 1.7Hz).H,Schematicrepresentationofthemicrofluidicdevice.I–K,Foldchangesinneuriteoutgrowth(lengthandbranches)upondifferentcombinationsofdoubleknockdown.AHPs weretransfectedwithtwodifferentconstructsencodedbyfluorescentcolors(mCherryandGFP)foreachofthecombinatorydoubleknockdownsetups(shSCR/scSCR,shSCR/shScg2,shREST/shSCR, and shREST/shScg2) and then cultured in the presence of FGF2 for 5 d. *p 0.05. **p 0.005. ns, Not significant.

Article Snippet: Recombinant SCG2 (MW 70.8 kDa, NP_003460) was purchased from Origene and used at 20 ng/ml, resuspended in 0.1% BSA in PBS.

Techniques: Recombinant, Staining, Cell Culture

Journal: Journal of Neuroscience

Article Title: REST Regulates Non-Cell-Autonomous Neuronal Differentiation and Maturation of Neural Progenitor Cells via Secretogranin II

doi: 10.1523/jneurosci.4286-14.2015

Figure Lengend Snippet: Figure6. SCG2isnecessaryandmediatesthenon–cell-autonomouseffectsofRESTknockdown.A,Inthetopchamberofthemicrofluidicdevice(treatmentchamber),conditionedmediumwas added; WT AHPs were plated in the bottom chamber (cell chamber). B, Anti-SCG2, but not control antibodies, can deplete the differentiation-inducing activity in shREST-conditioned medium. Conditionedmediumfromcontrolantibodyenhanceddifferentiation.C,AnincreaseinSox2(**p0.001)andadecreaseinTUJ1(***p0.0001)expressionwereobserveduponimmunodeple- tion (n 24). D, Average length analysis of neurites in immunodepletion experiments. A significant decrease in neurite length (***p 0.0001) was observed upon SCG2 immunodepletion. E, Sub-branches of neurites are observed only in SCG2 non-immunodepleted samples.

Article Snippet: Recombinant SCG2 (MW 70.8 kDa, NP_003460) was purchased from Origene and used at 20 ng/ml, resuspended in 0.1% BSA in PBS.

Techniques: Control, Activity Assay, Immunodepletion

Journal: iScience

Article Title: Highly efficient generation of self-renewing trophoblast from human pluripotent stem cells

doi: 10.1016/j.isci.2024.110874

Figure Lengend Snippet:

Article Snippet: Anti-CGA Antibody , Atlas Antibodies , HPA029698.

Techniques: Purification, Produced, Virus, Control, Recombinant, Lysis, Extraction, Cell Culture, Gene Expression, Protease Inhibitor, Bicinchoninic Acid Protein Assay, Western Blot, Library Quantification, Multiplex Assay, Sequencing, Software, Modification

Journal: Cell stem cell

Article Title: Formation of Human Neuroblastoma in Mouse-Human Neural Crest Chimeras

doi: 10.1016/j.stem.2020.02.001

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: For immunostainings, samples were blocked with 2% BSA and incubated with primary antibodies including Rabbit anti-ALK (1:200, Cell Signaling technology (CST), 3633), Rabbit anti-MYCN (1:50 84406; 1:100 51705, CST), Rabbit anti-CD3 (1:300, RM9107S0, Thermo Fisher), Rabbit anti-CD8a (1:300, 98941, CST), Rabbit anti-FoxP3 (1:75, MAB8214, R&D), Mouse anti-Il2ra (CD25, 1:100, NB600–564, Novus), Rabbit anti mouse Tim-3 (1:200, 83882, CST), Rabbit anti-human-PD-L1 (1:300, 13684, CST), Sheep anti-human-CD47 (1:100, AF4670SP, R&D), Mouse anti- IFNγ (1:200, DB-1, BioLegend), Rat anti-mouse F4/80 (1:100, MF48000, Thermo Fisher), Rabbit anti-γH2AX (1:300, ab2893, Abcam), Rabbit anti-human-Ki67 (1:20, PA516785, Thermo Fisher), Rabbit anti-Chromogranin A (1:500, NB120–15160, Novus), Rabbit anti-Synaptophysin (1:200, 36406, CST), Mouse anti-human-Nestin (1:300, ab176571, Abcam), Rabbit anti-TH (1:500, {"type":"entrez-protein","attrs":{"text":"P40101","term_id":"731386","term_text":"P40101"}} P40101 –150, Pel-Freez), Rabbit anti-pERK1/2 (1:400, 4370, SCT), Rabbit anti-p-p70 S6 (1:300, 9205, CST), Rabbit anti-Peripherin (1:500, ab4666, Abcam), Rabbit anti-human neurofilament (1:100, 160kD, ab92539, Abcam), anti-eGFP (1:1000, GFP-1020 Aves Labs; 1:300 AF647-Alpaca nanobody; 1:5000, ab290, Abcam), mouse anti-HNK-1 (1:50, C6680, MilliporeSigma) and rabbit anti-p75 (1:175, G3231, Promega) overnight at 4°C followed by appropriate secondary antibody incubation for 1–2 h (for IF: A31571, A21235, A21202, A11001, A21206, A11008, {"type":"entrez-protein","attrs":{"text":"A31573","term_id":"87384","term_text":"pir||A31573"}} A31573 , A21244, A11015 or A21209, Thermo Fisher; 703–545-155, 115–585-044, Jax ILR; for IHC: Rabbit-on-Rodent and Mouse-on-Mouse HRP-Polymer, RMR622; MM620, Biocare, or Biotinylated anti-rabbit IgG, BA-1000 Vector).

Techniques: Plasmid Preparation, Microarray, Recombinant, Isolation, SYBR Green Assay, Sequencing, Software