PIM-1 dependent VDR-transactivation. (A) PIM-1 overexpression enhances calcitriol-induced extrachromosomal DR3 reporter response. The luminescence signal increased after induction of the transfected HaCaT cells with 0.2 μM calcitriol for 16 h but not following the addition of an equal amount of ethanol for the same time. The cells transfected with PIM-1 showed a 2.8-fold increase in response to calcitriol compared with the cells transfected with empty vector (pDEST26/neg control). Also, similar signal intensities were observed for vehicle-treated cells transfected with PIM-1 vs. vehicle-treated cells transfected with the empty vector. (B) PIM-1 knockdown through shRNA plasmid constructs reduces the calcitriol-induced extrachromosomal DR3 reporter response to the half compared with the cells transfected with the neg. control shRNA plasmid. Furthermore, about the similar decrease was observed by calcitriol-independent extrachromosomal DR3 reporter response. Vehicle-treated cells transfected with shRNA against PIM-1 vs. vehicle-treated cells transfected with the empty vector (pGFP-V-RS/neg. control) reduces the signal intensity. Error bars represent the SEM of n = 3 values.