Background
Methods
Cell culture conditions and treatment
Western blot
Migration and invasion assays
Quantitative reverse transcriptase-PCR (qRT-PCR)
Transcript | Forward primer (5′-3′) | Reverse primer (5′-3′) |
---|---|---|
ACTB
| ATCGTCCACCGCAAATGCTTCTA | AGCCATGCCAATCTCATCTTGTT |
CDH1
| CCAGAAACGGAGGCCTGAT | CTGGGACTCCACCTACAGAAAGTT |
CTNND1
| ATGTTTGCGAGGAAGCCGC | CGAGTGGTCCCATCATCTG |
FOXC2
| GCCTAAGGACCTGGTGAAGC | TTGACGAAGCACTCGTTGAG |
Small-interfering (si) RNA-mediated knockdown
Gene | Sequences |
---|---|
PRKCA (ON-TARGET plus SMARTpool) | UAAGGAACCACAAGCAGUA UUAUAGGGAUCUGAAGUUA GAAGGGUUCUCGUAUGUCA UCACUGCUCUAUGGACUUA |
FOXC2 (ON-TARGET plus SMARTpool) | CCUACGACUGCACGAAAUA CCAACGUGCGGGAGAUGUU GGAUUGAGAACUCGACCCU GCGCCUAAGGACCUGGUGA |
FOXC2 (Dicer-substrates) | #1 5′ CGACUGCACGAAAUACUGACGUGTC 3′ 5′ GACACGUCAGUAUUUCGUGCAGUCGUA 3′ #2 5′ GGUGGUGAUCAAGAGCGAGGCGGCG 3′ 5′ CGCCGCCUCGCUCUUGAUCACCACCUU 3′ #3 5′ ACAUCAUGACCCUGCGAACGUCGCC 3′ 5′ GGCGACGUUCGCAGGGUCAUGAUGUUC 3’ |
Luciferase reporter activity assay
Confocal microscopy
Chromatin Immunoprecipitation (ChIP)
Forward primer (5′-3′) | Reverse primer (5′-3′) | |
---|---|---|
p120 ChIP (+127 to +309) | GATCCCGAAAGGAGGAAGAG | CGACTTGCTTATCCTCCTTTTCCC |
p120 non-specific 1 (−34 to +126) | GTACTTTGGCGGGGGAGATT | AGCAGGGCTGAAACCGATAC |
p120 non-specific 2 (+407 to +481) | GGCTGACATCACTTAGGAAAGC | CTCTTCCTCCTTTCGGGATC |