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[Preprint]. 2023 Jul 3:2023.07.03.547547. [Version 1] doi: 10.1101/2023.07.03.547547

Extended Data Fig. 2 |. β-hydroxybutyrate directly induces protein insolubility without posttranslational modification.

Extended Data Fig. 2 |

a, SDS-PAGE and quantification of Bis-ANS fluorescence in soluble fraction of native (50 kD, +37°C) and heat-misfolded (aggregated in well, +70°C) BSA treated with 10 mM of R-βHB and 10 mM of 1,3-BD. b, Western blot and quantification for KβHB in native (+37°C) and heat-misfolded (+70°C) BSA which remains soluble or becomes insolubilized following exposure to 5–10 mM of R-βHB and 1–2 mM of R-βHB-SNAC. c-e, Imperial staining and quantification of BSA which remains soluble or is insolubilized following (c) simultaneous exposure of BSA to 5–10 mM of R-βHB and 0.5–2 mM R-βHB-SNAC at +70°C, (d) native folded (+37°C) KβHB-BSA treated with 5–10 mM of R-βHB, and (e) heat-misfolded (+70°C) KβHB-BSA treated with 5–10 mM of R-βHB.

a, Representative image from triplicate repetitions.

b, Mean ± S.E.M, N=3, p-value calculated using one-way ANOVA with Sidak’s multiple comparison test.

c, Mean ± S.E.M, N=3, p-value calculated using one-way ANOVA with Dunnett’s multiple comparison test.

d, Mean ± S.E.M, N=3, p-value calculated using one-way ANOVA with Dunnett’s multiple comparison test.

e, Mean ± S.E.M, N=3, p-value calculated using one-way ANOVA with Dunnett’s multiple comparison test.