Fig 2: The knockdown of LRSAM1 inhibits autophagy and the CHE-induced rejuvenation in old HDFs. (A,B) Generation of old HDFs with LRSAM1 knockdown using three different of small hairpin RNAs that target LRSAM1. (C,D) Depletion of LRSAM1 suppresses lysosomal function induced by CHE. Old HDFs with LRSAM1 knockdown were treated with 20 μg/mL of CHE for 24 h. The cells were stained with lysotracker for 1 h, and the fluorescence intensity was quantified using image J software, * p < 0.05. (E,F) LRSAM1 is an important regulator for CHE-induced rejuvenation in old HDFs. Old HDFs with LRSAM1 knockdown were treated with 20 μg/mL of CHE for 24 h. The cells were stained with SA-β-gal solution and observed using microscopy. The results were normalized to the total cell number. Scale bar: 100 μm, ** p < 0.01.