Figure 5

Multiple Thap1 siRNA and shRNA sequences decrease level of the transfection- or transduction-dependent mouse and human 32 kDa T1-LIR in HEK293T cells. (a) HEK293T cells were transfected with pCDNA-hThap1 alone or combined with siRNA sequences specific to hThap1 (siRNA 1-hThap1), an unrelated cDNA sequence (GL2Luc), and/or an shRNA targeting GAPDH. Total cellular extracts (15 µg) were immunoblotted with Proteintech anti-Thap1 (top panel) or NeuroMab anti-Thap1 (bottom panel). (b) HEK293T cells were transduced with LV-mThap1 alone or together with shRNA sequences, #21 and #96, directed at mouse Thap1, or an shRNA non-silencing control (NS). Total cellular extracts (15 µg) were immunoblotted with Proteintech (top) or NeuroMab (bottom) anti-Thap1 antibodies. (c) Levels of 32 kDa Thap1 silencing in Proteintech and Neuromab immunoblots from (b), normalized to GAPDH. Control = untreated cells. Each graph is representative of 2 independent experiments.