Gut granules in <i>glo-3(kx38)</i> mutants.

(A-D) Signals from antibodies to PGP-2, RAB-7, LMP-1 and ectopically expressed CDF-2::GFP were acquired in 1.5-fold stage embryos using confocal microscopy. (A-B) In wild type and glo-3(kx38) mutants, CDF-2::GFP extensively colocalized with PGP-2 (white arrows in insets), however CDF-2::GFP was also mislocalized to the plasma membrane in glo-3(kx38) mutants (black arrow in B). (C-D) While PGP-2 compartments contained LMP-1 in wild type, the anti-LMP-1 signals were weak or lacking in glo-3(kx38) mutants (white arrows in insets) and often were located on the plasma membrane (black arrow in D). (A-D) Single optical sections are shown and black arrowheads flank the intestine. (E) SQUASSH software was used to calculate the area of PGP-2 organelles that contained the indicated marker in 5–7 embryos of each genotype. The mean is plotted, bars represent the 95% confidence intervals, * indicates p≤0.05, and ** indicates p≤0.001 by one way ANOVA. (F) Autofluorescent organelles were imaged in living pretzel stage embryos using wide field fluorescence microscopy. The diameters were determined for 25 randomly chosen compartments in six wild-type embryos and all of the compartments (ranging from 9–22 organelles) in six glo-3(kx38) mutants. (G) Confocal Z-stacks that span the entire intestine of 1.5-fold stage embryos stained with anti-PGP-2 antibodies were acquired. SQUASSH software was used to quantify the number of PGP-2 compartments in 5 embryos of each genotype. (E-G) In each graph the mean is plotted, bars represent the 95% confidence intervals, * indicates p≤0.05, and ** indicates p≤0.001 by one way ANOVA comparing glo-3(kx38) to wild type.