Fig. 6

Fig. 6 Embryonic exposure to AFB1 impairs gastrointestinal tract development in zebrafish larvae. (A) Heat map illustrating significant down-regulation of digestive tract development-associated gene set in the AFB1 (0.1 mg/L) -exposed 120 hpf zebrafish larvae compared to the control. Data represent the normalized gene expression values of the four independent biological replicates per treatment. Red asterisks indicate the selected genes for RT-qPCR. (B) RT-qPCR-based analysis of 2 selected gastrointestinal tract development-related genes after embryonic exposure to different sub-lethal concentrations of AFB1 (n = 5 replicates, n = 30 pooled larvae per replicate). (C) Length of the larval gut after AFB1 exposure (n = 3 replicates, n = 10 larvae per replicate). (D) Microsphere swallowing assay. Representative images of 120 hpf zebrafish larvae incubated in fluorescence microsphere suspension after AFB1 exposure. Scale bar = 500 µm (E) Scatter plot representing the integrated fluorescence intensities of the swallowed microspheres accumulated in the larval gut after AFB1 exposure (n = 2 replicates, n = 10 larvae per replicate). (F) Proportion of the larvae observed in different fluorescence categories indicating the microsphere accumulation level in the pharyngeal region (n = 2 replicates, n = 10 larvae per replicate). Fluorescence level was categorized as “low” < 5 × 104, 5 × 104 < ”medium”< 5 × 105 and 5 × 105 < ”high” (expressed as the sum of the pixel values). Data represent the mean and SD. “*” indicates statistical significance at p < 0.05 vs. the control (DMSO).