Fig 1: Rgs9 gene expression difference by the intracranial alternating current stimulation (iACS) treatments. Volcano plot of gene expression changes of neurons between sham and ips‐iACS groups in the cortex (A), hippocampus (E), and thalamus (I). Significant genes were called via DESeq2 (p < 0.05 and change >2‐fold). Uniform manifold approximation and projection (UMAP) showed Rgs9 expression in the cortex (B), hippocampus (F), and thalamus (J). Distribution and cell numbers of identified Rgs9 + neurons for sham and ips‐iACS groups in the cortex (C), hippocampus (G), and thalamus (K). The mRNA expression of Rgs9 in the iACS‐treated cortex (D), hippocampus (H), and thalamus (L) by real‐time PCR. The mRNA expression of Rgs9 by real‐time PCR were shown as mean ± standard deviation (SD), *** p < 0.001, ** p < 0.01, and *** p < 0.05 were considered as significantly different between each iACS and sham.
Fig 2: The immunofluorescence of Rgs9+ neurons in intracranial alternating current stimulation (iACS)‐treated rat brain. The NeuN+/RGS9 cells in the cortex (A), hippocampus (D), and thalamus (G). Quantification of RGS9+ cell count percentage in the cortex (B), hippocampus (E), and thalamus (H). Quantification of RGS9‐NeuN+/NeuN+ cell count percentage in the cortex (C), hippocampus (F), and thalamus (I). The nuclei were labeled with DAPI staining. The percentages of RGS9+/DAPI+, RGS9‐NeuN+/NeuN+ were shown as mean ± standard deviation (SD), *** p < 0.001, ** p < 0.01, and * p < 0.05 were considered as significantly different between each of the three iACS groups and sham. n = 3 rats for each group. Score bars: 200 or 100 µm as shown in each image.
Fig 3: Signaling pathway analysis of intracranial alternating current stimulation (iACS) regulation on neurons. The pathway enrichment analysis of down streaming upregulated genes and pathways in the Rgs9 + neurons from the cortex (A), hippocampus (B), and thalamus (C), using Gene Ontology (GO) biological process terms. (D) The protein expressions of RGS9, R7BP, Gβ5, and β‐catenin in the rat cortex with iACS treatments. (E) Quantification of RGS9 expression in (D). (F) Quantification of R7BP expression in (D). (G) Quantification of Gβ5 expression in (D). (H) Quantification of β‐catenin expression in (D). The protein expressions were shown as mean ± standard deviation (SD), *** p < 0.001, ** p < 0.01, and * p < 0.05 were considered as significantly different between each of the three iACS groups and sham. n = 3 rats for each group.
Supplier Page from Abcam for Anti-RGS9 antibody [EPR2873]