The sporadic sAD2.1 iPSNs increase the levels of OTULIN and phosphorylated tau, whereas inhibition of OTULIN deubiquitinase or deletion of OTULIN gene decreases phosphorylated tau levels. (IMAGE)
Caption
The sporadic sAD2.1 iPSNs increase the levels of OTULIN and phosphorylated tau, whereas inhibition of OTULIN deubiquitinase or deletion of OTULIN gene decreases phosphorylated tau levels. (A, B) OTULIN and phosphorylated tau levels were elevated in sporadic sAD2.1 iPSNs. Western blot analyses show significantly elevated phosphorylated tau at p-S199/p-S202/p-T205 (AT8), p-T231 (AT180), and p-S396/p-S404 (PHF-1) and OTULIN in sAD2.1 as compared to healthy controlWTC11 iPSNs. (C) A Connolly surface representation of the major atoms of UC495 that interact with the OTULIN deubiquitinase binding pocket are shown in the figure. The OUTLIN surface is shown with electrostatic potentials, with red color indicating regions of high negative potential (electron-rich), blue color indicating regions of high positive potential (electron-poor), and other colors representing intermediate potentials. (D, E) Pharmacological inhibition of OTULIN deubiquitinase with compound UC495 selectively decreases phosphorylated tau at AT8 site in sAD2.1 iPSNs. Western blot analyses showed significantly decreased AT8+ tau and amodest reduction of PHF-1+ tau upon inhibition of OTULIN deubiquitinase with compound UC495 in sAD2.1 iPSNs. (F, G) CRISPR-Cas9–mediated knockout (KO) of OTULIN causes a complete loss of total tau in sporadic sAD2.1 iPSNs.Western blot quantifications showing significantly reduced AT8, AT180, and PHF-1 positive as well as total tau in CRISPR-Cas9–mediated OTULIN KO of sporadic sAD2.1 as compared to sAD2.1 iPSNs. Note that the reduction is not due to toxicity to neurons, as no significant difference in the NeuN levels was noted.
Credit
Kiran Bhaskar
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