ANXII expression analysis by flow cytometry and cloned ANXII analysis by Western blotting. (A) Initial gating on total cell population. (B) Overlay histogram analyzing ANXII expression (in green) compared with isotype control (in red). (C) Graph illustrating the Kolmogorov-Smirnov test to calculate the D value (the green line is ANXII; the black line is the isotype control). The same strategy was used to analyze ANXII expression in CD117⁺ cells (D). (E) Western blot to determine ANXII expression in Pichia methanolica. Nine clones were tested for expression, and human serum albumin cloned into P methanolica (lane H) was used as the control (86 kDa). The approximate weight of the ANXII and pMET plasmid backbone was 41 kDa (lane 8).