Fig. 4: Loss of Pngl is associated with gut barrier defect which contributes to the lethality. | Nature Communications

Fig. 4: Loss of Pngl is associated with gut barrier defect which contributes to the lethality.

From: Gut barrier defects, intestinal immune hyperactivation and enhanced lipid catabolism drive lethality in NGLY1-deficient Drosophila

Fig. 4

a Low magnification fluorescent images of the midguts of indicated genotypes upon FITC-labeled dextran (yellow) feeding. b Confocal images showing phalloidin (magenta) and DAPI (blue) staining in the indicated genotypes upon FITC-labeled dextran (yellow) feeding. c Graph showing the quantification of gut barrier defect phenotype in the indicated genotypes (n = 60 for each genotype from three independent experiments). d Confocal images showing phalloidin (magenta), DAPI (blue), and HPA-lectin (yellow) staining for peritrophic matrix in control and Pngl mutant midguts. e Light microscopic images of cross-section of control and Pngl mutant midguts. L, lumen; PM, peritrophic matrix; PS, peritrophic space; EC, enterocytes. f Graph showing quantification of gut barrier defect phenotype in the indicated genotypes (n = 60 for each genotype from three independent experiments). g Graph showing % lethality rescue in the indicated genotypes. h Confocal images showing Phalloidin (magenta), DAPI (blue), and ConA (yellow) lectin staining in the proventriculus region of third instar larvae of indicated genotypes (n = 6–7 for each genotype). Insets show close-up of PM-secreting cells. i Graph showing the quantification of the number of intracellular puncta in the indicated genotypes. j Graph showing the size of intracellular puncta in the indicated genotypes (n = 12–30 puncta from three animals per genotype). k Confocal images showing Phalloidin (magenta), DAPI (blue), and ConA (yellow) lectin staining in the proventriculus region of third instar larvae of indicated genotypes (n = 4 for each genotype). Insets show a close-up of PM-secreting cells. l Graph showing the quantification of the number of intracellular puncta in the indicated genotypes. m Graph showing the size of intracellular puncta in the indicated genotypes (n = 9–14 puncta from four animals per genotype). The scale bar is 250 µm in (a) and 50 µm in (b, d, h, k). In all panels, each circle represents an independent replicate in (c, f, g, i) or a punctum in (j, m) and mean ± standard deviation is shown. P values are indicated on bars. Significance is ascribed as P < 0.05 using one-way ANOVA followed by Šidák correction. Source data are provided as Source Data file.

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