Journal: Journal of cell science
Article Title: STIM1L traps and gates Orai1 channels without remodeling the cortical ER.
doi: 10.1242/jcs.164228
Figure Lengend Snippet: Fig. 5. Cluster expansion requires the STIM1 lysine-rich tail but not cytosolic Ca2+ elevations. (A) Left, TIRF images of cells expressing YFP–STIM1 and YFP–STIM1L taken before (left) and 10 min after thapsigargin (Tg) addition (right) in conditions preventing cytosolic Ca2+ elevations (20 mM BAPTA-AM and 50 mM La3+). Insets, a 2.6-fold magnification to show the morphology of fluorescent clusters. Scale bars: 10 mm. Right, quantitative analysis of TIRF images showing the effects of thapsigargin on the absolute increase in cluster size. (n533/5/3, 38/5/3, 36/5/3 and 44/5/3 cells/recordings/transfections). (B) Left, TIRF images of cells expressing YFP–STIM1DK and YFP–STIM1LDK taken before (left) and 10 min after thapsigargin addition (right). Right, quantitative analysis of TIRF images showing the effects of thapsigargin on the absolute increase in cluster size. (n541/7/6, 49/6/5, 51/7/5 and 42/6/5 cells/recordings/transfections). Quantitative data show the mean6s.e.m.; *P,0.05, **P,0.01; ****P,0.0001; ns, not significant (unpaired Student’s t-test).
Article Snippet: YFP– STIM1DK was obtained from Addgene (Plasmid 18861; Cambridge, MA) and YFP–STIM1LDK was obtained by mutagenesis (GeneCust; Dudelange, Luxembourg), Orai1 was obtained from Addgene (Plasmid 12199; Cambridge, MA) and pCMV/myc/ER/GFP (KDEL–GFP) was purchased from Life Technologies (Plasmid V823-20).
Techniques: Expressing, Transfection