Fig. 2
From: Optogenetic protein clustering through fluorescent protein tagging and extension of CRY2
Enhancement of light-induced CRY2 clustering by a C-terminal extension. a Sequences of CRY2 variants extended at the C-terminus of CRY2PHR [a.a. 1–498] with the nine residues of a new peptide (blue box) or those of native CRY2 [a.a. 1–507]. Mutated residues are shown in red. b Fluorescence images of a cell showing reversible clustering of mCherry-CRY2clust. Bottom, kymograph corresponding to the yellow line in the top left image. c Graph of time-lapse results showing changes in cluster ratio. Both yellow and blue arrows in b, c indicate illumination time points. d Fluorescence images of cells expressing each of the indicated mCherry-labeled CRY2 variants, upon light stimulation. Cells were illuminated with light for 5 min at 20 s intervals. e Quantification of cluster ratio. Values are expressed as means ± s.e.m. (n = 72, 99, 91, 76, 98, 88, 97 cells, three independent experiments). **P = 3.45 × 10–8; ***P = 5.57 × 10–14, ****P = 9.42 × 10–62 (CRY2PHR), 1.32 × 10–69 (CRY2clust(L7K)), 9.97 × 10–70 (CRY2clust(L7D)) by Student’s two-tailed t-test. Scale bars, 20 μm
