Journal: bioRxiv

Article Title: Dynamic S-acylation of STIM1 is required for store-operated Ca 2+ entry

doi: 10.1101/2022.02.15.480522

Figure Lengend Snippet: A) Schematic representation of the inactive and active (extended) conformation of STIM1. The locations of the EF hand, coiled-coil 1 (CC1) and STIM1 Orai1 activation region (SOAR) of STIM1 are indicated. The location of cysteine 437 is noted at the C-terminus of the SOAR domain. For simplicity, one monomer of the dimeric STIM1 structure is presented. B) Jurkat T cells were treated with anti-CD3 antibody for 0, 1, 2, 5, 10, and 15 minutes and subjected to acyl-RAC. The reaction without hydroxylamine (-HA) serves as a negative control. Calnexin serves both as a positive control for S-acylation and a loading control. Representative of n=5. C) Quantification of fold change of S-acylation of STIM1 normalized to calnexin. Error bars indicate S.D. D) Input blots are shown for levels of pPLCγ1, PLCγ1, STIM1, and calnexin. The pPLCγ1 blot is indicative of the time course of TCR activation. E) HEK STIM double knockout (DKO) cells were transfected with WT or C437S STIM1-FLAG and acyl-RAC was performed after 5 minutes of calcium store depletion with thapsigargin. The blots are representative of four independent experiments.

Article Snippet: For immunoblotting, the antibodies were purchased from commercially available sources: STIM1 (#4961S), calnexin (#2679), phospholipase Cγ1 (PLCγ1) (#2822S), phospho-PLCγ1 (Tyr783) (#2821S), anti-rabbit IgG, HRP-linked secondary antibody (#7074S), anti-mouse IgG, HRP-linked secondary antibody (#7076P2) were from Cell Signaling Technology; anti-CD3 human antibody (14-0037-82) was from eBiosciences.

Techniques: Activation Assay, Negative Control, Positive Control, Control, Double Knockout, Transfection

Journal: Nature neuroscience

Article Title: Psychedelics promote plasticity by directly binding to BDNF receptor TrkB.

doi: 10.1038/s41593-023-01316-5

Figure Lengend Snippet: Fig. 5 | Psychedelics recruit neurotrophic signaling through BDNF and TrkB. a, Representative single-molecule localization microscopy maps of dendritic spines after treatment with vehicle, LSD and PSI. TrkB in light blue, PLCγ1 in yellow, TrkB:PLCγ1 in dark blue, PLCγ1:TrkB in red. Scale bar, 200 nm. b,c, Psychedelics increase colocalization of TrkB and PLCγ1 in dendritic spines. d, Timeline of LSD-induced increase in pERK. e, M100907 pretreatment does

Article Snippet: Materials & experimental systems n/a Involved in the study Antibodies Eukaryotic cell lines Palaeontology and archaeology Animals and other organisms Human research participants Clinical data Dual use research of concern Methods n/a Involved in the study ChIP-seq Flow cytometry MRI-based neuroimaging Antibodies Antibodies used anti-TRKB (R&D systems, AF1494); https://www.rndsystems.com/products/human-mouse-rat-trkb-antibody_af1494 anti-PLCγ1 (Cell Signaling Technologies, 5690S); https://www.cellsignal.com/products/primary-antibodies/plcg1-d9h10-xp-rabbitmab/5690 anti-phospho-TrkB(Tyr816) (Cell Signaling Technology, 4168S); https://www.cellsignal.com/products/primary-antibodies/phosphotrka-tyr785-trkb-tyr816-c67c8-rabbit-mab/4168 anti-TRK (Cell Signaling Technologies, 92991); https://www.cellsignal.com/products/primary-antibodies/trk-pan-a7h6r-rabbitmab/92991 anti-phospho-mTOR (Cell Signaling Technology, 5536S); https://www.cellsignal.com/products/primary-antibodies/phospho-mtorser2448-d9c2-xp-rabbit-mab/5536 anti-phospho-p44/42 MAPK (Erk1/2) (Cell Signaling Technology, 9106S); https://www.cellsignal.com/products/primary-antibodies/ phospho-p44-42-mapk-erk1-2-thr202-tyr204-e10-mouse-mab/9106 anti-mTOR (Cell Signaling Technology, 2972S); https://www.cellsignal.com/products/primary-antibodies/mtor-antibody/2972 anti-p44/42 MAPK (Erk1/2) (Cell Signaling Technology, 9102S); https://www.cellsignal.com/products/primary-antibodies/p44-42mapk-erk1-2-antibody/9102 anti-MAP2 (Invitrogen, PA5-17646); https://www.thermofisher.com/antibody/product/MAP2-Antibody-Polyclonal/PA5-17646 anti-MAP2 (Abcam, ab5392); https://www.abcam.com/map2-antibody-ab5392.html anti-BrdU (Abcam, ab6326); https://www.abcam.com/brdu-antibody-bu175-icr1-proliferation-marker-ab6326.html 3 nature portfolio | reporting sum m ary M arch 2021 anti-NeuN (Millipore, MAB377, clone A60); anti-β-actin (Sigma-Aldrich, A1978); Alexa Fluor 488 donkey anti-rat (Invitrogen, A21208); Alexa Fluor 647-conjugated antibody goat anti-chicken (Invitrogen, A-21449); Alexa Fluor 647-conjugated donkey anti-goat (Invitrogen, A32849); Alexa Fluor 568 goat-anti-mouse (Invitrogen, A11004); CF568-conjugated donkey anti-rabbit (Biotium, 20098); HRP-conjugated GaR (BioRad, 170-5046); HRP-conjugated RaG (Invitrogen, 61-1620); HRP-conjugated GaM (Bio-Rad, 170-6515) Validation All these antibodies were validated by the manufacturer and used in previous studies from our laboratory (e.g. Casarotto, P. C. et al. Antidepressant drugs act by directly binding to TRKB neurotrophin receptors.

Techniques: Microscopy

Journal: Cell stem cell

Article Title: Human iPSC-derived Natural Killer Cells Engineered with Chimeric Antigen Receptors Enhance Anti-Tumor Activity

doi: 10.1016/j.stem.2018.06.002

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit monoclonal anti-phospho-PLCγ1 (Ser1248) , cell signaling technology , Cat#8713 RRID: AB_10890863.

Techniques: Virus, Expressing, Plasmid Preparation, Recombinant, Transduction, Apoptosis Assay, Cell Isolation, Membrane, Protein Extraction, Enzyme-linked Immunosorbent Assay, Sequencing, Software, Imaging

Journal: Cell Reports

Article Title: CDK8 Fine-Tunes IL-6 Transcriptional Activities by Limiting STAT3 Resident Time at the Gene Loci

doi: 10.1016/j.celrep.2020.108545

Figure Lengend Snippet:

Article Snippet: Mouse anti-human-PLCγ1-AF647 (Clone 27/PLC) , BD Biosciences , Cat#557883; RRID: AB_396921.

Techniques: Purification, Recombinant, Software