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Fig. 3 | Acta Neuropathologica Communications

Fig. 3

From: Altered ribosomal function and protein synthesis caused by tau

Fig. 3

7 days of hTau and FTD-mutant hTau expression alter RP abundance, protein synthesis and ribosomal biogenesis. a 7 days of hTau overexpression decreases protein synthesis and alters the abundance of select ribosomal proteins (RPs) in HEK293 cells. Cells expressing EGFP, hTau-EGFP, P301L-hTau-EGFP or K369I-hTau-EGFP were selected by using neomycin. Newly synthesised proteins were labelled for 16 h with AHA, before collection at 7 days post transfection. FUNCAT-WB showed that protein synthesis was decreased by hTau expression, with this decreasing being greater in FTD-mutant hTau expressing cells. hTau, P301L-hTau, and K369I-hTau expression also lead to decreased abundance of RPL5 and RPS14. The abundance of RPS6 was increased in P301L-hTau expression cells compared to EGFP controls. Protein abundance was normalised to the total protein stain REVERT. n = 3 wells, one-way ANOVA, Tukey’s MCT. b Polysome, monosome and 60S ribosomal subunit abundance is decreased by 7 days of hTau expression in HEK293 cells. Transfected cells were treated with 100 ug/mL CHX for 5 min in order to prevent the dissociation of bound ribosomes from mRNA and following lysis, ribosomal complexes were separated on a 10–50% linear sucrose gradient via ultracentrifugation. 40S and 60S ribosomal subunit, along with monosome and polysomes, were detected via their absorbance at 260 nm, with the area under the curve (AUC) of these peaks being used for quantification. The abundance of the 60S ribosomal subunit, monosomes and polysomes was decreased in all three hTau expressing groups. n = 3 wells, one-way ANOVA, Tukey’s MCT

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