Fig. 3: N-acetylcysteine reduces L68Q-hCC density in sucrose-gradient ultracentrifugation.
From: NAC blocks Cystatin C amyloid complex aggregation in a cell system and in skin of HCCAA patients
L68Q-hCC cell lysates were treated with β-mercaptoethanol (β-MetOH) or n-acetyl cysteine (NAC) for 1 h at 37 °C. Lysates were layered on top of sucrose step gradients, ranging from 5% sucrose at the top (fraction 1) to 30% at the bottom (fraction 9) in 5% increments. Gradients were ultracentifuged in a swinging bucket rotor for 16 h at 4 °C and 246,000×g. Fractions were collected from gradients, proteins were precipitated with tricholoroacetic acid, and precipitated proteins were dissolved in PBS before analysis by SDS-PAGE under reducing conditions. Gels were blotted for cystatin C. The image is representative of three independent experiments and sucrose gradients.
