Fig. 4

Fusion with a large fluorescent protein, tandem dimeric Tomato, further decreases CRY2low homo-oligomerization. Blue light was delivered at 5-s intervals for 100 s (a, b) or at 2-s intervals for 20 s (c, d). a In cells expressing both CRY2wt-GFP-Sec61 and CIB1-tdTom, blue light induced fewer clusters formation compared to cells expressing only CRY2wt-GFP-Sec61. b Quantification of the cluster mass of CRY2wt-GFP-Sec61, with or without co-expressing CIB1 fusion proteins, shows that CIB1-tdTom suppresses CRY2 cluster formation. (n = 11, 16, 11, 10). c After recruitment to the ER membrane via CRY2-CIB1 hetero-dimerization, CRY2wt-tdTom formed fewer clusters than CRY2wt-mCh, and CRY2low-tdTom did not form visible clusters on the ER network. d Illustration of CRY2 homo-oligomerization suppressed by the large protein tdTom. e The cluster mass of CRY2 on ER membrane after recruitment via CRY2–CIB1 hetero-dimerization was quantified and normalized against that of CRY2wt-mCh. (n = 16, 12, 15, 14). Results are presented as means ± s.e.m. and analyzed using one-way ANOVA with Dunnett’s post hoc test. (**P < 0.005, ***P < 0.0005). Scale bars, 5 µm