Introduction
Method
Human studies
DNA methylation study
Eye measurements performed at 3 years old
Animal studies
FDM mice and measure of ocular axial length
Cellular studies
Human retinal pigment epithelial cell culture
Mouse primary RPE cell
Immunocytochemistry for mouse RPE cells
miRNA transfection
Construction and transfection of HOXA9 cDNA
Cell viability assay
EdU proliferation assay
Real-time PCR, western blot
Statistical analysis
Results
Human methylation study for pre-school children
Increased HOXA9 expression in myopic eyes in the FDM animals
Gene | Primer |
---|---|
HOXA9 F | 5′- CCC CGA CTT CAG TCC TTG C- 3’ |
HOXA9 R | 5′- GAT GCA CGT AGG GGT GGT G- 3’ |
PAX6 F | 5′- TAG CCC AGT ATA AAC GGG AGT G- 3’ |
PAX6 R | 5′- CCA GGT TGC GAA GAA CTC TG- 3’ |
MMP2 F | 5′- CAA GTT CCC CGG CGA TGT C- 3’ |
MMP9 R | 5′- TTC TGG TCA AGG TCA CCT GTC-3’ |
FGF2 F | 5′- GCG ACC CAC ACG TCA AAC TA-3 ‘ |
FGF2 R | 5′- TCC CTT GAT AGA CAC AAC TCC TC-3 ‘ |
IGF1R F | 5′- GAA GAA CGC CGA CCT CTG TTA- 3’ |
IGF1R R | 5′- GCA GCG ATT TGT GGT CCA G ‘ |
GAPDH F | 5′- TGA CCA CAG TCC ATG CCA TC-3 ‘ |
GAPDH R | 5′- GAC GGA CAC ATT GGG GGT AG-3 ‘ |