Figure 5. Effects of AMPK knock-down with shAMPKα1 on SA-galactosidase expression.

a. Lentivirus vectors expressing shRNA targeting human AMPK alpha subunit were created. One of them knocked-down the AMPK alpha1 subunit more than 95%, 48 hours after infection, without affecting alpha2 expression. Total AMPK alpha was suppressed more than 80%. b, c. Keratinocytes grown in 12 well plates were infected with lentivirus expressing control (non-targeting) shRNA (shControl) or AMPK alpha1 (shAMPKα1) 48 hours prior to incubating for 2 hours with H₂O₂±1 mM AICAR SA-Gal activity was assessed 64 hours later. In fection of the cells with shAMPKα1 both significantly increased basal SA-Gal (*, p<0.05 vs shControl+0 mM H₂O₂, n = 4) and eliminated the preventive effects of AICAR (†, p<0.05 vs shAMPKα1+0 mM H₂O₂, n = 4).