Dynamic AFM imaging of HCQ-induced formation of formation of AnxA5 layer over aPL IgG-β₂GPI complexes. Each original AFM amplitude image (top panel) is paired with a colorized version (bottom panel) for explanation. In a sequence performed without HCQ, (A) the addition of β₂GPI to phospholipid bilayers resulted in the formation of clusters (marked 1-4, orange circles in colorized version), as previously described.⁴¹  (B) Addition of AnxA5 resulted in the initiation of 2-dimensional crystallization (flat round structures, green in colorized version) arising from different nucleation points, which (C) enlarge, merge, and grow toward each other, as previously described,⁷  until they covered all of the bilayer not occupied by the β₂GPI cluster. (D) Subsequently added aPL mAb bound to the surfaces of β₂GPI clusters. In a sequence that included HCQ, (E) addition of β₂GPI to phospholipid bilayers and formation of β₂GPI clusters (marked 1-5) were followed by (F) addition of HCQ, followed by (G) addition of AnxA5, which at first crystallized from multiple different nucleation sites on the phospholipid bilayer, (H) and then formed a secondary layer of AnxA5 over the β₂GPI, which progressively grew around and over the adjacent primary layer of AnxA5 crystal, leaving none of the surface uncovered by AnxA5.