Extended Data Fig. 6: CsLinker expression recovers pyrenoid formation in ΔEPYC1 Chlamydomonas.
From: A promiscuous mechanism to phase separate eukaryotic carbon fixation in the green lineage
a, Example of Rubisco partitioning calculation using integrated density analysis in Fiji, according to ref. 15. b, Rubisco partitioning in the condensate in the WT (CrRbcS-mCherry/EPYC1-Venus), ΔEPYC1 (CrRbcS-mCherry) and ΔEPYC1 (CrRbcS-mCherry/CsLinker) lines, quantified from the images in e and Supplementary Fig. 15 using the method outlined in a. Statistical significance from unpaired t-tests are indicated; **** = p < 0.0001. c, Area of the Rubisco condensate as measured in Fiji, according to region ‘A’ in a. For the ΔEPYC1 (CrRbcS-mCherry), the largest condensed fluorescence signal at the canonical position was measured. *** = p < 0.001. The median and quartile values are represented by the solid and dashed horizontal lines in each plot. d, Estimated volume distributions of the Rubisco condensates in the three lines, assuming sphericity of the condensate and calculating from the cross-sectional area in b. *** = p < 0.001. e, Confocal fluorescence microscopy images of tagged RbcS and CsLinker in the ΔEPYC1 background. Scale bars = 1 μm. f, Western blot confirmation of CsLinker variant expression in WT and ΔEPYC1 background lines. g, Western blot confirmation of CsLinker expression in ΔEPYC1 background relative to the empty vector and background strains. RbcL was used as a loading control.
