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. 2020 Aug 6;31(1):103–119. doi: 10.1111/bpa.12883

Figure 4.

Figure 4

Overexpression of active PKR increases tau phosphorylation and tau expression. A. Immunoblot of whole‐cell lysates from HEK 293T cells using total‐ and pTau‐specific antibodies as well as anti‐V5 tag to detect V5‐tagged constitutively active D328A PKR. Cells were co‐transfected with vectors expressing 2N4R tau and TD Tomato or 2N4R tau and V5‐tagged constitutively active D328A PKR for 24 h. Cells were treated with GSK3i or DMSO as vehicle control for the last 4 h prior to cell lysis. Figure is representative of five independent experiments. B. Quantifications of total tau/α‐tubulin from five independent experiments (n = 5, **P < 0.01, student t‐test). C. Quantifications of the effect of GSK3 inhibition on different phospho‐tau epitopes/total tau in cells co‐expressing 2N4R tau and V5‐tagged constitutively active D328A PKR. Results from three independent experiments (n = 3, *P < 0.05, student t‐test). D. mRNA levels in RNA extracts were quantified by qPCR. PKR mRNA levels are normalized to NADH mRNA. The values in the figure exhibit the ratio between cells expressing tau and TD Tomato and cells expressing tau and D328A PKR (n = 4, **P < 0.01, student t‐test). E. mRNA levels in RNA extracts were quantified by qPCR. Tau mRNA levels are normalized to NADH mRNA. The values in the figure exhibit the ratio between cell expressing tau and TD Tomato and cells expressing tau and D328A PKR (n = 4, **P < 0.01, student t‐test).