HDAC1 Silencing Regulates CIITA Expression

The PMA treated differentiated THP-1 cells (10⁵ cells/ml) were transiently transfected with 600 pmole (Garcia-Garcia et al., 2009 (link)) of siGENOME Human HDAC1 (3065) siRNA – SMARTpool (Dharmacon, USA) using lipofectamine 3000. The cells were incubated with siRNA for 24 h to allow gene silencing. Subsequently, the cells were washed for infection and stimulated as described earlier. THP-1 cells were harvested after 6 h and mRNA expression of HDAC1 and CIITA genes was analyzed by qPCR. ON-TARGET plus Control Pool (Dharmacon, USA) was used as a negative control. The basal level of HDAC1 and CIITA in uninfected Sc-siRNA transfected cells respectively were used for data normalization and were taken as 1.0. The transfection efficiency was calculated (ThermoFisher) to be >50%, as has also been reported by the manufacturer (Dharmacon, USA).