Effect of KU-0063794 and KU-0068650 compounds compared to Rapamycin in in vitro and ex vivo keloid models. (a) Both KU-0063794 and KU-0068650 inhibit the expression of collagen, fibronectin, and α-smooth muscle actin (α-SMA) at messenger RNA (mRNA) levels (keloid fibroblast (KF): n=8). (b) In-Cell Western Blotting analysis of the expression of ECM proteins, Cyclin D, and proliferating cell nuclear antigen (PCNA), 24 hours post treatment with different mammalian target of rapamycin inhibitors (KF: n=8). (c) Shrinkage of keloid organ culture (OC) after different compound treatments. Average weight of the keloid OC (n=10) at different time points are indicated in the bar graph. Four millimeter keloid explant biopsies were removed from the collagen gel matrix and its weight was determined in triplicates. (d) In MTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay both, KU-0063794 and KU-0068650 showed much greater inhibitory effect on metabolic activity in keloid OC (n=8) as compared with Rapamycin. *P<0.05, **P⩽0.01 indicates significant difference compared with the DMSO group.