The dynamin-like protein Fzl promotes thylakoid fusion and resistance to light stress in <i>Chlamydomonas reinhardtii</i> - Fig 5

(a) Fluorescence microscopy of cells from wild-type (top) and a ΔCrFzl strain expressing the CrFzl-mVenus construct (bottom) after growth in normal (NL) and high light (HL) conditions. Chlorophyll autofluorescence is used as a thylakoid membranes marker. Scale bar: 1μm. (b) Subdomain organization of the distinct CrFzl-3HA versions expressed in the ΔCrFzl mutant background. (c-d) Western blot analysis of the expression of the distinct CrFzl-3HA versions expressed in the ΔCrFzl mutant with anti-Tubulin, anti-HA (c) and anti-CrFzl (d). Untransformed wild-type and ΔCrFzl strains are used as controls. (e) Spot assay of the parental wild-type, the ΔCrFzl strain #1, the strain ΔCrFzl #1 complemented with CrFzl-3HA and ΔCrFzl #1 expressing a mutant version of CrFzl-3HA (left panel: CrFzl-G1*-3HA; middle panel: CrFzl-ΔHD-3HA; right panel: CrFzl-ΔCC-3HA) on TAP at normal light (NL, 50 μmol.m-2.s-1) as a control, and on minimal TP medium at 100 μmol.m-2.s-1 (TP HL). (f) Western blot analysis of CrFzl-3HA, CrFzl-G1*-3HA and CrFzl-ΔHD-3HA in thylakoid fractions washed with 0 and 500 mM NaCl after 6 hours of normal (NL) or high light (HL) treatment. Loadings were adapted to reach equivalent amounts of β-CF1. 0 and 500 mM NaCl samples were loaded on the same gels; uncut western blots are available in S4c Fig. (g) Percentage of CrFzl-3HA, CrFzl-G1*-3HA or CrFzl-ΔHD-3HA left in thylakoid fractions washed with 500 mM NaCl after 6 hours of normal (NL) or high light (HL) treatment. HA signal was normalized to the β-CF1 signal and compared with the amount found in thylakoids washed without NaCl. Error bars represent the s.e.m. from three independent experiments.