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Fig. 6 Gene set enrichment analysis (GSEA) and qPCR validation reveal Tl-induced transcriptional changes related to neural and cardiovascular development, partially reversed by ER stress modulators. (A) Ridge plot based on RNA-seq analysis shows significantly enriched gene sets between the control and TI groups, with color gradients representing adjusted p-values. Nervous system development-related GO BP pathways include (B) axon development (GO), positive regulation of nervous system development (GO), cell morphogenesis involved in neuron differentiation (GO), and cardiovascular system-related pathways GO BP pathways include blood vessel morphogenesis (GO), heart morphogenesis (GO), and heart development (GO) were enriched in control group compare to TI. Each subplot visualizes the ranked distribution and enrichment score curves, highlighting the distribution of genes within the ranked gene sets. All the gene sets from (B) to (G) have normalized enrichment score (NES) < -1.5, p value < 0.001, and q value < 0.001. (H) natriuretic peptide B (nppb) and (I) jun B proto-oncogene (junb), associated with heart development, and (J) neuritin 1-like b (nrn1lb) and (K) neurotrophin 3 (ntf3), associated with neural development, were analyzed in the control group, Tl (800 μg/L) group, Tl + IXA4 (20 μM) group, and Tl + TUDCA (20 μM) group after exposure from 6 to 48 hpf. Data are presented as average fold changes relative to the control group, with n = 3 per group. Statistical significance was determined using unpaired t-test analysis. *p < 0.05, * *p < 0.01, * **p < 0.001, compared to the indicated group.