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. 2015 Jun 1;43(13):6285–6298. doi: 10.1093/nar/gkv576

Figure 3.

Figure 3.

Inhibitory effect of TLP on the processing of TFIIAαβ. TFIIAαβ protein was determined by Western blotting. (A) Effect of overexpressed TLP on intracellular TFIIAαβ protein. TLP- and N37E-introduced HCT116 cells were treated with CHX for the indicated time, and the amount of endogenous TFIIA protein was determined. (B) Effect of TLP knockdown on TFIIAαβ precursor. The amount of endogenous TFIIAαβ precursor in TLP-knocked-down HeLa cells was assayed as described above. (C) Effect of TLP on TFIIAαβ processing. HeLa cells were transfected with expression plasmids for FH-TLP and FH-N37E together with FH-TFIIAαβ, and the TFIIAαβ precursor and generating α subunit were detected. (D) Effect of TLP on the uncleavable DGAA mutant. HeLa cells were transfected with expression plasmids of FH-TFIIA and FH-DGAA mutant together with TLP, and the amount of the FH-TFIIAαβ precursor was determined. FH-MyoD introduced into cells was used as an electrophoresis loading standard. (E) RT-PCR for detection of TFIIA-L transcripts. TLP-overexpressed HCT116 cells were assayed for TFIIA-L transcripts by RT-PCR using specific primers.