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. 2019 Jun 13;8:e46300. doi: 10.7554/eLife.46300

Figure 5. Altered ALA-induced proteolysis of GluTR1 in FC2 knockout mutants.

(A) Levels of FC2 mRNA in light-grown Col-0 and fc2-2 plants, analyzed by qRT-PCR. Data are given as mean ± sd (n = 3). n.d., not detectable (B) Levels of GluTR1, FC1, ClpC and ClpS in leaves of Col-0 and fc2-2 plants incubated without (-) or with (+) 1 mM ALA in darkness (D) for the indicated times. Signal intensities for GluTR1 relative to Col-0 seedlings harvested before the beginning of the experiment (Col-0 t(0)) are shown (D, dark). (C) and (D) Levels of HEMA1 (C) and GSA1 (D) mRNAs in fc2-2 mutants plotted relative to untreated Col-0 seedlings harvested before start of the experiment. Data are given as mean ± sd (n = 4). Letters indicate statistical groups determined by Student’s t-test (p<0.05). (E) Protoporphyrin IX (ProtoIX), (F) Pchlide, (G) ncb heme and (H) free heme contents of Col-0 and fc2-2 leaves treated as in (B). The inset in E shows the steady-state level of ProtoIX in untreated seedlings before ALA treatment (t0). Data are given as mean ± sd (n = 4). Asterisks denote statistically significant changes between samples with p<0.05 (Student’s t-test). Statistical analysis was performed before the calculation of ratios. n.d., not detectable. 21-day-old, light-grown plants were analyzed.

Figure 5.

Figure 5—figure supplement 1. ALA-induced proteolysis of GluTR1 in FC1 knockdown mutants.

Figure 5—figure supplement 1.

(A) FC1 mRNA levels in leaves of light-grown Col-0 and fc1-1 plants, analyzed by qRT-PCR. Data are given as mean ± sd (n = 3). The asterisk denotes the significant reduction of FC1 transcript in the fc1-1 mutant (Student’s t-test, p<0.05). (B) Levels of GluTR1, GBP, ClpC and FC1 in leaves of Col-0 and fc1-1 plants incubated without (-) or with (+) 1 mM ALA in darkness for the indicated times. (C, D) Levels of Pchlide (C) and ncb heme (D) in leaves of Col-0 and fc1-1 plants treated as in (A). Data are given as mean ± sd (n = 4). Values were tested (Student’s t-test, p<0.05) for significant differences between Col-0 and fc1-1 at each time point. The mutant accumulated the same amounts of Pchlide and heme as the WT. n.s., not significant.