Journal: Molecular cell
Article Title: The Allergen Der p3 from House Dust Mite Stimulates Store-Operated Ca 2+ Channels and Mast Cell Migration through PAR4 Receptors.
doi: 10.1016/j.molcel.2018.03.025
Figure Lengend Snippet: Figure 4. Ca2+ Influx to Mite Extract Requires STIM1 Gating of Orai1 (A) Western blots compare STIM1 and Orai1 protein expression before and after knockdown (KD). (B) Aggregate data from two independent experiments are shown. (C) Mite-induced Ca2+ influx is reduced following knockdown of either STIM1 or Orai1. (D) Aggregate data from several experiments as in (C) are compared. HDM (400 mg/mL) is the mean of 159 cells, KD STIM1 126 cells, and KD Orai1 81 cells. (E) TIRF images of STIM1-YFP puncta are compared between resting HEK293 cells and cells stimulated with mite extract. Images for the stimulated condition were taken 9 min after mite exposure in Ca2+-free solution. (F) Confocal images compare STIM1-YFP distribution in HEK293 cells at rest, after stimulation with thapsigargin (8 min), and after exposure to mite extract (15 min). (G) The rate of Ca2+ entry, normalized to control (mock-transfected cells), is compared for the conditions shown. HDM bar denotes mean of 44 cells, STIM1 38 cells, and STIM1+Orai1 30 cells. (H) Aggregate data are compared. Orai1 was knocked down and then the various constructs indicated were expressed (together with STIM1) 24 hr later. GFP bar denotes mean of 66 cells (which were transfected only with GFP plasmid and served as control), - 52 cells, Orai1 33 cells, L273S-Orai1 37 cells, and 81AARAE85- Orai1 15 cells. For the graphs, all data are presented as mean ± SEM.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Anti-STIM1 Cell Signaling 5668 Anti-Orai1 Santa Cruz sc-68895 Anti-His Santa Cruz sc-8036 Bacterial and Virus Strains BL21(DE3), E. coli Lucigen 60401-2 Biological Samples Human nasal mast cells ENT Department N/A Chemicals, Peptides, and Recombinant Proteins Der p1 protein Indoor Biotechnologies RP-DP1D-1 Der p3 protein Bioclone PQ-0335 House dust mite Greer XPB70D3A2.5 Nafamostat mesylate Sigma N0289 Camostat mesylate Sigma SML0057 Aprotinin Sigma A1153 E-64 Sigma E3132
Techniques: Western Blot, Expressing, Knockdown, Control, Transfection, Construct, Plasmid Preparation
Cell Signaling Technology Inc is a verified supplier 
Cell Signaling Technology Inc manufactures this product 
