Fig 1.
Experimental design and differential gene expression workflow.
A) Delta-9-tetrahydrocannabinol dosing timeline for fetal and infant rhesus macaque cohorts. B) RNA sample processing and differential gene expression workflow using NanoString nCounter. Created with BioRender.com.
Fig 2.
In utero THC-exposure shifts gene expression at the fetal and infant timepoints.
A) Heatmaps of DEG between control and THC-exposed offspring in the fetal (left) and infant (right) cohorts. Values were normalized in ROSALIND by dividing gene counts within a lane by the geometric mean of normalization probes from the same lane. Differential gene expression was assessed using a generalized linear model, assuming a negative binomial distribution. B) PCA plots of control (red) vs THC-exposed (blue) differential gene expression. C) Contributions of DEG to differences between control and THC-exposed groups. The red line denotes variability contribution percentage if each gene contributed equally.
Table 1.
DEG in the fetal cohort.
Table 2.
DEG in the infant cohort.
Fig 3.
Gene enrichment analysis using STRING pathway and Enrichr.
A) Gene enrichment analysis using STRING to highlight the pathways most affected by in utero THC exposure in fetal (left) and infant (right) cohorts. Fetal DEG involved in PI3k-Akt signaling pathway are highlighted in red. Infant DEG involved in inflammatory response, IL-10 signaling, and IL-4 and IL-13 signaling are highlighted in red, blue, and green, respectively. B) Gene enrichment analysis using Enrichr. Bar charts of the top 10 enriched terms using the WikiPathway 2023 Human gene set library for fetal (top) and infant (bottom) cohorts.
Fig 4.
Chromatic staining of fetal and infant rhesus macaque rectus femoris tissue.
A) Representative images of Masson-Goldner Trichrome staining of fetal (top) and infant (bottom) rectus femoris tissues. Whole tissue scale bars measure 2000 μm, zoomed image scale bars measure 100 μm. B) Quantification of collagen in Masson-Goldner Trichrome stain in fetal and infant cohorts. Horizontal lines represent the mean of each group. Statistical analysis was performed using unpaired t-test on GraphPad Prism 10.