Figure 5.

Distribution of ChlH/GUN5 in Lysed and Fractionated Chloroplasts That Were Either Fed or Not fed with PPIX.

(A) Analysis of lysed and fractionated chloroplasts by immunoblotting with anti-ChlH/GUN5 antibodies. The same fractions described in Figure 4 were analyzed by immunoblotting with anti-ChlH/GUN5 antibodies. As described for Figure 4, 200 μg of purified intact chloroplasts were either fed (+) or not fed (−) with 20 μM PPIX. These chloroplasts were then fractionated into soluble (S) and membrane-containing pellet (P) fractions of equal volume. Equal volumes were analyzed by SDS-PAGE and immunoblotting.

(B) Proportions of ChlH/GUN5 in soluble and membrane fractions. The percentage of ChlH/GUN5 in the pellet (white bars) and supernatant (light-gray bars) fractions derived from chloroplasts that were not fed PPIX and the percentage of ChlH/GUN5 in the pellet (medium-gray bars) and supernatant (dark-gray bars) fractions derived from chloroplasts that were fed PPIX are indicated for the wild type (Col-0) and each mutant and transgenic line. Results from seven independent experiments are shown for Col-0, five independent experiments are shown for cs, and four independent experiments are shown for all other mutants and transgenic lines. Error bars indicate se. The asterisk indicates a statistically significant difference in the levels of ChlH/GUN5 in the membrane-containing pellet fractions between Col-0 and gun4-1 (P = 0.02), between Col-0 and cch (P = 0.008), and between gun5 (P = 0.04), gun5-101, and cs (P < 0.002).