Figure 5. Reduced sCLU membrane levels in hemolytic anemia correlate to markers of RBCs senescence, redox imbalance and erythrophagocytosis.

(A₁) Representative immunoblots of RBCs plasma membrane preparations from a healthy subject (Control-1) and two patients with hereditary spherocytosis (HS-2 and HS-4) exhibiting decreased sCLU membrane levels. (A₂) Representative immunoblot analysis of a RBCs membrane preparation from the splenectomized patient HS-1; sCLU membrane levels are similar to controls. Immunoblots were also probed with antibodies against Band 3, Hb, dinitrophenylhydrazone (DNP) residues (oxyblot analysis), CD47, Hsp70, IgGs and actin (used as loading control); in some cases compositions of different blots is shown. Molecular weight markers are indicated at the right of each blot. (B) Densitometric analyses of sCLU and cellular senescence or oxidative stress markers in respective immunoblots. Shown data are the mean values of the proteins' relative proportion against a reference membrane protein followed by normalization to the controls (N = 10, non-smoking, age-matched subjects). Error bars, ±s.d.; single or double asterisks, difference of patients vs. controls at significance level of p<0.05 or p<0.01, respectively.