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. 2020 Sep 15;94(19):e00420-20. doi: 10.1128/JVI.00420-20

FIG 6.

FIG 6

Mutation at the NS3-4A, but not the NS4A-4B or NS4B-5A, junction blocks NS5A hyperphosphorylation. (A) Schematics of NS3-5A expression constructs with mutations at the NS3-4A, NS4A-4B, or NS4B-5A cleavage junction. The sites of mutation are numbered and indicated with one-letter amino acid symbols: C, cysteine; D, aspartate; F, phenylalanine; R, arginine; S, serine; T, threonine; Y, tyrosine. (B to F) Representative immunoblotting for NS3, NS5A, and NS5A phosphorylated at S2201 (pS2201), S2208 (pS2208), S2211 (pS2211), and S2214 (pS2214) in HEK293T cells transfected with the constructs. (G) Changes (percent) in p58 over total NS5A (p56 plus p58). Values are means ± SEM (n = 3). *, P < 0.05 (determined by t test against the values for the wild type). (H) NS5A phosphorylation at S2201, S2208, S2211, and S2214 normalized with total NS5A and relative to values for the wild type. *, P < 0.05 (determined by t test against the values for the wild type).