Figure 4

A 50 kDa T1-LIR species is detected by Proteintech antibody in mouse neural tissue and testes and is developmentally regulated. (a) Nuclear (150 µg) and cytoplasmic (150 µg) extracts from adult heart, testis, and whole brain were subjected to SDS-PAGE and immunoblotted with Proteintech anti-Thap1 antibody. The 29 kDa T1-LIR species (*) was detected in both compartments in all three tissues. The 32 kDa species (**) was detected only in nuclear extract from testis. The 50 kDa species (arrow) was detected in the nuclear compartment in testis (doublet) and brain, with a trace amount detected in cytoplasm, likely due to contamination with nuclear proteins (not shown). (b) Total cellular extracts (50 µg) from adult peripheral organs (left panel) and adult and E15 individual brain regions (right panel) were immunoblotted with Proteintech anti-Thap1 antibody. “Positive control lysate” lane corresponds to HEK293T cells transfected with a human Thap1 expression vector. (c) Extracts from liver, cerebellum and forebrain from P1 mice were subjected to SDS-PAGE, immunoblotted, and patterns were compared to those observed in adult cerebellum. Total cellular RIPA (150–200 µg), cytoplasmic (cyt) (150–200 µg) and nuclear (nuc) extracts (50–75 µg) were immunoblotted with Proteintech antibody. The lower 50 kDa T1-LIR species (**) was detected only in brain tissues and the levels of the upper 50 kDa (arrow) and lower 50 kDa were increased in P1 relative to adult. Note that the 50+ kDa species in P1 liver nucleus does not co-migrate with either of the two brain species. The 29 kDa species (*) appeared in nuclear extract only in neural tissue.