DNMT1 H3 DNMT3b
PBS Scr ShRNA
mTOR GAPDH
PBS Scr ShRNA
0 0.5
1 1.5
DNMT3b DNMT1 mTOR
Relative level
Supplementary Fig. 1
Supplementary figure 1. Expression of DNMT3b, DNMT1, and mTOR proteins in the ipsilateral
L4/5 DRGs of naive rats 5 weeks after DRG microinjection of PBS, AAV5-Dnmt3a shRNA (ShRNA) or AAV5-scambled shRNA (Scr). Left: representative Western blots. Right: statistical summary of the densitometric analysis.
Supplementary Fig. 2
B
MOR KOR Tubulin DNMT3a H3
Relative level
GFP + SNL GFP + Sham Cre + SNL
0 0.5 1 1.5 2 2.5
**
MOR KOR
** **
##
##
DNMT3a
##
Cre + Sham GFP + Sham
Cre + SNL PBS + SNL GFP + SNL
A
Relative level (Ipsi/Con) ** ## **
Oprd1
##**
Oprk1
**
** ******
Oprm1 0
1 2 3 4
**
**
##
**
**
Dnmt3a
Sham
+ GFP GFP Cre SNL
Supplementary figure 2. DNMT3a participates in nerve injury-induced downregulation of MOR
and KOR expression in mouse injured DRG. Expression of Dnmt3a, Oprm1, Oprd1, and Oprk1 mRNAs in the ipsilateral (Ipsi) or contralateral (Con) L4 DRGs (A) and expression of DNMT3a, MOR, and KOR proteins in the ipsilateral L4 DRG (B) on day 7 after L4 SNL or sham surgery in the Dnmt3afl/fl mice. SNL or sham surgery was carried out 5 weeks after microinjection of PBS, AAV5-Cre (Cre), or AAV5-GFP (GFP) into unilateral L4 DRG in mice. n = 12 mice/group. **P
< 0.01 vs the corresponding sham plus GFP group. ##P < 0.01 vs the corresponding SNL plus GFP group.
Relative level
HSV-DNMT3a + AAV5-GFP HSV-GFP + AAV5-Cre
AAV5-Cre + HSV-DNMT3a Dnmt1 Dnmt3b Oprm1 Oprd1 Oprk1
**
**
**
**
## ##
PBS
HSV-GFP + AAV5-GFP
Dnmt3a
**
**
##
0 5 10 15 20 25
0 0.5
1 1.5
2 2.5
3 3.5
Supplementary Fig. 3
Supplementary figure 3. DNMT3a regulation of MOR and KOR expression in the cultured
mouse DRG neurons. Expression of Dnmt3a, Dnmt3b, Dnmt1, Oprm1, Oprd1, and Oprk1 mRNAs in the cultured DRG neurons of Dnmt3afl/fl mice. The cultured neurons were treated with PBS or transduced as indicated.
Supplementary Fig. 4
WT KO
MBD1 H3
WT HET KO
350 bp 450 bp
A B
Supplementary figure 4. Representative tail genotyping from Wild type (WT), heterozygous (HET), and Mbd1 knockout (KO) mice (A) and representative Western blots from the DRGs of wildtype and Mbd1 KO mice (B).
