Figure 1
From: Optogenetic induction of caspase-8 mediated apoptosis by employing Arabidopsis cryptochrome 2
Design of optogenetic tools to control cell apoptosis by blue light. (A) Blue light induces CRY2-PHR-mcherry oligomerization in HEK293T cells. a. dark; b. blue light 1 min; c. blue light 5 min; d. blue light 10 min; e. blue to dark 5 min; e. blue to dark 10 min. (B) The photo-responsive region of Arabidopsis thaliana cryptochrome 2 (PHR, amino acid 1–498) is fused with Caspase8. In the dark, the engineered PHR-Casp8 exhibits monomer. Upon blue light illumination, PHR drives dimerization of PHR-Casp8 and promoters the apoptosis. Casp8, Caspase8; Casp3, Caspase3; PHR, the N-terminal ___domain of CRY2. (C) Clustering of caspase-8-PHR-mCherry in response to blue light in HEK293T cells, bar = 10 μm. (D) Blue light actives Opto-Caspase8-V1 in HEK293 T cells. Transfected HEK293T cells were crushed by Pierce IP lysis buffer, total cell lysates were analyzed by western blotting probed with anti-Caspase8, anti-Caspase3, actin was used as a loading control.
