To down-regulate miR-203 in HaCaT cells, we used an antisense miR-203 vector (MZIP203-PA-1) and a miRZip scrambled hairpin control vector (MZIP000-PA-1) from System Biosciences (SBI; CA, USA). HaCaT cells were transfected with the pZip control and pZip-203 vectors using the Effectene reagent (Qiagen; MD, USA). Three days after the transfection, transfected cells were enriched by sorting based on the copGFP reporter, followed by puromycin selection. Two pooled miR-203 knockdown cell clones (Zip203-1 and Zip203-2) were generated via independent transfection followed by the selection procedures described above.
We used an miR-146a-overexpressing vector (PMIRH146aPA-2) and a scrambled control hairpin in pCDH-CMV-MCS-EF1-copGFP (CD511B-1) from SBI to generate stable miR-146a-overexpressing and control cell clones, respectively, using a similar protocol as describe above.
We used an miR-146a-overexpressing vector (PMIRH146aPA-2) and a scrambled control hairpin in pCDH-CMV-MCS-EF1-copGFP (CD511B-1) from SBI to generate stable miR-146a-overexpressing and control cell clones, respectively, using a similar protocol as describe above.