Fig. 6: Hypoxia-driven pathomechanism profiling in motor assembloids and a transcriptomics-guided drug discovery strategy. | Nature Communications

Fig. 6: Hypoxia-driven pathomechanism profiling in motor assembloids and a transcriptomics-guided drug discovery strategy.

From: Geometrically-engineered human motor assembloids-on-a-chip for neuromuscular interaction readout and hypoxia-driven disease modeling

Fig. 6

a Volcano plot of differential gene expression in whole motor assembloids from IH (n = 5 assembloids) and Ctrl (n = 4 assembloids). b Enrichment of KEGG pathways in whole motor assembloids, focusing on key pathways involved in environmental information processing and metabolism. c Heatmap showing the relative changes in gene expression within motor assembloid subfields, based on GSEA analysis of GO terms related to neuromuscular synaptic transmission. d Heatmap presenting the relative changes in gene expression of MN vulnerability and resistance markers within motor assembloid subfields, with red triangles indicating significance. e Schematic outline of the pathological target identification and drug discovery strategy. f Enrichment map and cluster analysis in whole motor assembloids. The network map visualizes the top 150 GSEA GO terms, with GO terms represented as nodes that are connected and clustered into functional themes based on shared gene associations. The functional theme of oxidative phosphorylation (OXPHOS) is highlighted, with the left plot showing all associated GO terms within this cluster. g Representative immunoblots (left) and heatmap profiles (right) showing the protein levels of mitochondrial respiratory chain complex subunits in motor assembloids exposed to IH, with Ponceau S staining (total protein) as loading control. n = 6 independent experiments. h Enzyme activity of mitochondrial complexes I–V in motor assembloids across Ctrl and IH groups. n = 3 independent experiments. i and j Representative image (i) of ROS intensity in hMNS exposed to IH, with quantification (j) of ROS distribution radially across the spheroids and along the longitudinal diameter of hSkM. k Schematic of NAD+ conversion and consumption in the NAD+ salvage pathway. l–n Quantification of NAD+, NADH levels, and NAD+/NADH ratio in motor assembloids. n = 3 independent experiments. o Quantification of ATP level in motor assembloids. n = 3 independent experiments. Data are presented as mean ± SD. Statistical analysis: g, two-tailed Student’s t test. h, two-tailed Student’s t test and Mann-Whitney test. l–o One-way ANOVA with Bonferroni multiple comparison. ***p  <  0.001, **p <  0.01, *p  <  0.05. Source data are provided as a Source Data file.

Back to article page