( A) Confirmation of viral expression was performed in the hippocampus of 12-week-old mice, 14 days after stereotaxic delivery of lentivirus. Cryosections were stained for turbo GFP (tGFP) and DAPI. Note top panels denote side injected with the virus (‘Ipsi’, ipsilateral) and bottom panels, the uninjected side (‘Contra’, contralateral). (B) Confirmation of Trim28 knockdown was performed as in A, but using an antibody directed against Trim28. (C) The effect of Trim28 knockdown on α-Syn and tau in primary neurons can be rescued by overexpressing an shRNA-resistant cDNA of TRIM28, (D) TRIM28 knockdown in human cells does not alter MAPT transcript levels and only mildly affects SNCA expression as assayed by qRT-PCR. Trim28 heterozygous (Trim28+/-) mice have normal levels of Snca and Mapt in their brain. (E) TRIM28 knockdown does not alter protein levels of several other neurodegenerative disease-causing genes. HEK293T cells were infected with virus targeting TRIM28 (shTRIM28-1) or control (non-silencing, shScram) and levels of neurodegenerative disease-causing genes were measured by Western blot and quantified below. Data are presented as mean + s.e.m. In C, n = 3–4 replicates, *, ** and *** denote p<0.05, p<0.01 and p<0.001, respectively, One-Way ANOVA followed Holm-Sidak post-hoc test; in D, n = 3 replicates per group, top panel: *** and ns denote p<0.001 and p>0.05, respectively, One-Way ANOVA followed by Holm-Sidak post-hoc test, bottom panel: * and ns denote p<0.05 and p>0.05, respectively, Student’s t-test; in e, n = 3 replicates per group, ns denotes p>0.05, Student’s t-test.