Inactivation of Prions and Amyloid Seeds with Hypochlorous Acid
ScBH was pretreated for 1 h in BrioHOCl at 100:1 v/v BrioHOCl to 10% BH (panel A, blue), 20:1 BrioHOCl (panel B, blue) or corresponding saline treatments (panels A and B, red). Similar BrioHOCl and saline pretreatments of NBH are indicated in gray. Resulting samples were then subjected to serial 10-fold dilutions and RT-QuIC analysis was performed with hamster (90–231) rPrPC substrate using 2 μl per well of the indicated tissue dilutions as reaction seeds. The dilutions noted refer to the final dilution of original brain mass used to seed the reaction. The orange dashed line indicates the 50-h cutoff time at which seeding activity was quantified by end-point dilution under these assay conditions, after which unseeded control reactions occasionally gave false-positive reactions (see Material and Methods). Each trace represents the average ThT fluorescence of 4 technical replicate wells normalized between baseline and maximal signal and graphed as a function of time.