Olig1. Re la tiv e m. RN. A le v e ls. (A. U) h i g f. CON. GF. exGF. Figure S2: mRNA expression levels of myelin component and upstream myelin regulatoryĀ ...
Total RNA Extraction Pooling n=4 per group Randomized
cDNA synthesis
Sample Pooling n=12 per group
Illumina RNA Sequencing
CON
GF
PFC qRT-PCR validations males n=12 per group
Randomized pooling of male PFC samples
RNA Sequencing n=4 per group
CON
n=3 per group males only
xGF
GF
Animal sacrifice and perfusion
Total Protein extraction/concentraion determined
Post fixation sectioning and imaging
Minimum 50 axons per animal measured
Western Blot and analysis
g-ratio analysis
Animal Cohort 2
Additional female cohort n=12 per group Sfig.2
Brain region dissection (6 regions per animal)
Figure 3 . TEM
Cohort 2
n=7 per group randomly selected
Functional annotation of differentially expressed genes (GO-Terms & KEGG pathways)
12 animals per group male only
qRT-PCR- based expression analysis for myelin component and regulatory genes
Figure 4. Western Blot
Differential gene expression analysis
Animal Cohort 1
xGF
GF
CON
Other brain regions for qRT-PCR analysis males only n=12 per group
PFC dissection males only n=3 per group
TEM g-ratio analysis
Randomized selection of PFC samples for Protein extraction males only
Western Blot n=7 per group
Figure S1: Experimental flowchart. Graphic al depiction of the animal number and usage for each individual experiment. Each Panel represents the outcome of tissue used for each of the individual main figure.
Figure S2: mRNA expression levels of myelin component and upstream myelin regulatory genes in the PFC of female CON, GF and exGF mice. No male-equiv alent changes in myelin component mRNA levels were found in the PFC of female GF and exGF mic e. (a-i) qRT-PCR of myelin component gene transcript and myelin regulating transcription factors in the PFC. Bar graphs indicate average values of 12 animals after š¯›½ -actin normalization relative to average control levels (p0.05; n=3 animals per group; n=>50 axons per animal (CON n=187 axons; GF n=390 axons).
