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Figure 2 | Acta Neuropathologica Communications

Figure 2

From: Dystonia type 6 gene product Thap1: identification of a 50 kDa DNA-binding species in neuronal nuclear fractions

Figure 2

Primary striatal neurons contain endogenous 32 and 50 kDa T1-LIR species in the nuclear compartment, and the 32 kDa species increases following transduction with Ad-hThap1. (a) Total cellular extract (50 µg) from untreated (control) mouse striatal neurons blotted with the Proteintech antibody reveals endogenous 29, 50 and 75 kDa T1-LIR species. Following transduction with Ad-hThap1, a transduction-dependent 47 kDa species is detected (**), the 50 kDa species increases slightly in intensity (arrow) and a prominent 32 kDa species (*) appears, as detected by the Proteintech and NeuroMab antibodies. (b) An aliquot (40 µg) of protein from the cytoplasmic compartment from adult mouse brain and untreated and Ad-hThap1-transduced primary striatal neurons was separated and immunoblotted as indicated. The 29 kDa T1-LIR species was prominent in all 3 samples, whereas the 50 kDa (arrow) was detected only in primary striatal neuronal extract. T1-LIR species of greater apparent Mr were present in all 3 samples. (c) The 29 (*), 50 doublet (brackets) and 75 kDa (arrow) T1-LIR species were detected in 10 µg of adult brain nuclear extract, whereas, in primary striatal neuronal extract, a 32 kDa T1-LIR (**) was detected and the upper band of the 50 kDa T1-LIR species was relatively more prominent (brackets). Note that the NeuroMab and Santa Cruz antibodies do not detect all the identical species, but all recognize the 32 kDa species (**), which was observed to increase following transduction with Ad-hThap. The transduction-dependent 47 kDa species is recognized only by Proteintech (empty arrow). SC3H3 and NeuroMab antibodies also recognize a 27 kDa T1-LIR species in adult brain (filled arrowhead) and a very prominent 50+ kDa species (empty arrowhead).

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