(A) Hierarchical cluster dendrogram with colors underneath denoting
three distinct modules of co-expressed transcripts in RiboTag mice RNA-seq data.
(B) Heat map showing module-trait relationships, each row representing a module.
Correlation coefficients between a Module Eigengene (ME) and trait (coded from
−1 to 1) and corresponding P values are shown in each
cell. The color indicates the level of correlation (positive correlation in red
and negative correlation in green). Individual plots showing the expression of
the module Eigengene across sample categories with significantly enriched terms
for biological processes revealed by gene ontology enrichment analysis (FDR
< 0.05). (C) Cluster dendrogram identified 16 co-expressed WGCNA modules
for RiboTag AAV RNA-seq data. (D) Module-trait relationships revealed modules
correlated with hPMCA2w/b expression in striatal astrocytes. Green module was
composed of genes regulating protein transport, which were down-regulated in
striatal astrocytes expressing hPMCA2w/b. Pink module was enriched in genes
regulating chromatin silencing and transcription, which were upregulated in
striatal astrocytes expressing hPMCA2w/b. (E) Expression levels of GABA
transporters: Slc6a1 (GAT-1), Slc6a13 (GAT-2),
Slc6a11 (GAT-3) and Slc6a12 (GAT-4) in
RiboTag mice and RiboTag AAV RNA-seq data sets. (F) Expression levels of
potential GABA transporter regulators suggested by previous work in RiboTag mice
and RiboTag AAV RNA-seq data. (G) Changes in gene expression of striatal
astrocytes expressing hPMCA2w/b, expressed as log2 (fold-change), in
RiboTag mice and RiboTag AAV RNA-seq data. Asterisk (*) indicates differential
expression between hPMCA2w/b IP and control IP using edgeR analysis on combined
RNA-seq datasets (FDR < 0.1). (H-I) Cartoon summary of the main findings
at synaptic (H) and in vivo levels (I). (J) Descriptive model
based on our work (see discussion). Average data shown as mean ± SEM from
4 mice in each group.