h1 male human embryonic stem cell line  (WiCell Research Institute Inc)

Journal: Cell

Article Title: Neuromodulator Signaling Bidirectionally Controls Vesicle Numbers in Human Synapses

doi: 10.1016/j.cell.2019.09.011

Figure Lengend Snippet: Highlights:

Article Snippet: Male human embryonic stem cells (ESCs) line H1 (WA01 (WiCell Research Resources WiCell, WI) were purchased from WiCell (normal karyotype, passage 40) and used for all stem-cell-derived human neurons experiments.

Techniques: Transduction, Recombinant, Plasmid Preparation, Expressing, Software

Journal: Cell

Article Title: Neuromodulator Signaling Bidirectionally Controls Vesicle Numbers in Human Synapses

doi: 10.1016/j.cell.2019.09.011

Figure Lengend Snippet: Highlights:

Article Snippet: EXPERIMENTAL MODEL AND SUBJECT DETAILS Human Embryonic Stem Cells Male human embryonic stem cells (ESCs) line H1 (WA01 (WiCell Research Resources WiCell, WI) were purchased from WiCell (normal karyotype, passage 40) and used for all stem-cell-derived human neurons experiments.

Techniques: Transduction, Recombinant, Plasmid Preparation, Expressing, Software

Journal: Cell

Article Title: Neuromodulator Signaling Bidirectionally Controls Vesicle Numbers in Human Synapses

doi: 10.1016/j.cell.2019.09.011

Figure Lengend Snippet: Highlights:

Article Snippet: Human Embryonic Stem Cells Male human embryonic stem cells (ESCs) line H1 (WA01 (WiCell Research Resources WiCell, WI) were purchased from WiCell (normal karyotype, passage 40) and used for all stem-cell-derived human neurons experiments.

Techniques: Transduction, Recombinant, Plasmid Preparation, Expressing, Software

Journal: Neuron

Article Title: Neuroligin-4 regulates excitatory synaptic transmission in human neurons

doi: 10.1016/j.neuron.2019.05.043

Figure Lengend Snippet: (A) Representative images of patch clamping of EGFP labeled AD-iN cells in mixed cultures. (B) Quantification of intrinsic properties of spiked-in AD-iN cells in mixed cultures differentiated from wild-type H1 (WT), NLGN4KO (KO) and NLGN4R704C (R704C) cells. (C) Example mPSCs traces recorded in the presence of TTX, (D) quantification of mIPSCs amplitudes and frequencies, (E) quantification of mEPSCs amplitudes and frequencies from spiked-in AD-iN cells WT, KO and R704C at day42. (F) Example traces and quantification of sEPSCs and (G) evoked EPSC amplitudes recorded from AM-iN cells WT, KO and R704C cells at day42. (H) Representative images of surface (live staining) and total HA signal (fixed staining) in day 28 Ngn2 iN cells expressing NGLN4-HA or NLGN4R704C-HA. (I) Quantification of surface localized NLGN4 relative to total NLGN4 in Ngn2 iN cells (left) expressing NGLN4-HA (WT) or NLGN4R704C-HA (R704C). Right panel, immunoblotting from the same cultures shows comparable levels of total NLGN4 protein in WT and R704C. (J) NLGN4 co-immunoprecipitates with GluA1 and PSD-95. The R704C mutation enhances co-immunoprecipitation of NLGN4 with GluA1. Protein lysates from Ngn2 iN cells expressing NLGN4 WT, R704C or EGFP (Ctrl) immunoprecipitated with HA antibodies and blotted for the AMPAR-GluA1 and for PSD-95. Left, quantification of the relative levels of GluA1 and PSD95 in the immunoprecipitates. Scale bars: 50μm for panel A and upper panel H and 10μm lower panel H. Data are represented as mean ± SEM and N=3. Numbers of neurons/ independent cultures analyzed are shown in the bars. (*, p < 0.05) (***, p < 0.001). See also Figure S4.

Article Snippet: Male human embryonic stem (ES) cells line H1 (WA01 WiCell Research Institute, Inc.) were cultured up to passage 29 and authenticated by GTW banding karyotype method, only cells with normal 46, XY karyotype were used for experiments.

Techniques: Labeling, Staining, Expressing, Western Blot, Mutagenesis, Immunoprecipitation

Journal: Neuron

Article Title: Neuroligin-4 regulates excitatory synaptic transmission in human neurons

doi: 10.1016/j.neuron.2019.05.043

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Male human embryonic stem (ES) cells line H1 (WA01 WiCell Research Institute, Inc.) were cultured up to passage 29 and authenticated by GTW banding karyotype method, only cells with normal 46, XY karyotype were used for experiments.

Techniques: Recombinant, Reverse Transcription Polymerase Chain Reaction, Sequencing, Software