Journal: Cell reports

Article Title: Variation of Human Neural Stem Cells Generating Organizer States In Vitro before Committing to Cortical Excitatory or Inhibitory Neuronal Fates

doi: 10.1016/j.celrep.2020.107599

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: The following primary antibodies were used at the concentration indicated by manufacturer’s protocol: CaM Kinase II alpha (6G9) (NB100–1983), LMX1A (NBP1–81303) Novusbio; SYNAPSIN (106 001), HOMER (160 003) Synaptic System; EGFR (Ab231), FGFR1 phosphoY654 (Ab59194), TBR1 (Ab31940), REELIN (Ab18570), CYCLIN D1 (Ab10540), FGFR2 (Ab10648), BMPR1A (Ab38560) Abcam; HES1 (11988), p-SMAD1/5 (9516), CYCLIN D1 (2926), pERK1/2 (4370), FGFR1 (9740) Cell Signaling Technology; PAX6 (PRB-278P) BioLegend; NESTIN (MAB1259), OTX2 (AF1979), PDGFR alpha (AF1062; AF307), SOX2 (AF2018; MAB2018), SOX21 (AF3538), TuJ1 (MAB1195), EGFR (AF1280), O4 (MAB 1326) R&D Systems; GFAP (Z 0334) DAKO; HES5 (sc-13859), CUX1 (sc-13024), TLE4 (sc-9125), FGFR3 (sc-9007), LHX2 (sc-19344) Santa Cruz Biotechnology; FOXP2 (ABE73), TBR1 (AB2261), REELIN (MAB5366) Millipore; FOXG1 (M227) Takara/Clontech; anti GAD65/67 was kindly gifted by Dr. Christian Geis, Hans Berger Department of Neurology, Jena University Hospital, Germany ( ).

Techniques: Virus, Plasmid Preparation, Recombinant, Transfection, Antibody Labeling, In Vitro, Microarray, Gene Expression, Derivative Assay, Software

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: START: A Versatile Platform for Bacterial Ligand Sensing with Programmable Performances.

doi: 10.1002/advs.202402029

Figure Lengend Snippet: Figure 7. Design and characterization of STARTs for tetracycline and MS2 coat protein. a) Detailed schematics of the tetracycline Apta-trigger, Tetra_B3. The lower stem region of tetracycline aptamer subject to engineering is marked by a gray box. b) GFP fluorescence of toehold switch B and trigger RNA pairs in the absence (gray bars) and presence (green bars) of 12 μм tetracycline. Positive control (PC) used the toehold switch trigger B without the aptamer sequence, while negative control (NC) used the decoy RNA without any predicted interactions. c) GFP fluorescence output for switch B and Tetra_B3 pair for different concentrations of tetracycline inputs. d) Detailed schematics of the MS2 coat protein Apta-trigger, MS2_C1. The lower stem region of MS2 aptamer subject to engineering is marked by a gray box. e) GFP fluorescence of toehold switch C and trigger RNA pairs without (gray bars) and with (green bars) MS2 induction, by 10 ng mL−1 anhydrotetracycline (aTc) treatment. PC used the toehold switch trigger C, and NC used the decoy RNA. f) GFP fluorescence output for switch C and MS2_C1 pair for different induction levels for MS2 expression. P-values were determined by an unpaired t-test, with P > 0.05 designated by “ns”, P ≤0.05 designated by “*”, and P ≤0.0001 designated by “****”. Error bars are the SD from three biological replicates.

Article Snippet: The tetM expression plasmid for tetracycline resistance[64] was purchased from Addgene (pMflT-o4, #101312), and the plasmid was used without further modification.

Techniques: Positive Control, Sequencing, Negative Control, Expressing

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: START: A Versatile Platform for Bacterial Ligand Sensing with Programmable Performances.

doi: 10.1002/advs.202402029

Figure Lengend Snippet: Figure 8. Orthogonality assessment between STARTs. a) Experimental scheme to test the orthogonality of switch and theophylline Apta-trigger pairs. b) Crosstalk measured by flow cytometry for all switch-trigger combinations. Relative fluorescence was determined by taking the GFP output measured for a given trigger-switch combination and dividing it by the GFP output measured for the switch with its cognate Apta-trigger (diagonal). Theophylline was treated at a concentration of 10 mм. c) Experimental scheme to test orthogonality for different ligand inputs. d) Crosstalk measured by flow cytometry for all switch-trigger pairs and input ligands. Relative fluorescence was determined by taking the GFP output for a given Apta-trigger-switch pair and ligand combination and dividing it by the GFP output measured for the Apta-trigger-switch pair with its cognate input ligand (diagonal). Theophylline was treated at 10 mм, tetracycline was treated at 12 μм, and aTc was treated at 10 ng mL−1. Relative fluorescence value for each condition represents the mean of three biological replicates.

Article Snippet: The tetM expression plasmid for tetracycline resistance[64] was purchased from Addgene (pMflT-o4, #101312), and the plasmid was used without further modification.

Techniques: Cytometry, Concentration Assay