Fig. 6
a Expression levels by RT-qPCR of PHB1 and PHB2 in media supplemented with different concentrations of ammonium ferric citrate, measured by RT-qPCR. The dark-grey columns correspond to the PHB1 expression levels. The light-grey columns correspond to the PHB2 expression levels. The bars in the graphs represent the mean ± the standard deviation (SD). b Purification of PHB1 and PHB 2 from L. major homogenate by affinity chromatography using a Fe-NTA column. Identification by Western blot using anti-Leish r PHB1 (Lane 1) and Leish r PHB2 antibodies (Lane 2). Lane 3: native PHB1 eluted from the Fe-NTA column and stained with K3[Fe(CN)6III]and (4) Leish r PHB2 also stained with K3[Fe(CN)6III]. 2. *** P < 0.001 by the Tukey-Kramer multiple comparisons statistical test. c Protective effect of PHB1 and PHB2 against DNA damage caused by hydroxyl radicals. The positions to which supercoiled (I), relaxed circular (II) and linear (III) monomeric DNAs migrated are indicated on the left of the picture. Lanes 2, 3 and 4: native PHB1 at 500, 250 and 125 μg/ml, respectively; Lanes 5, 6 and 7: recombinant Leish r PHB2 at 500, 250 and 125 μg/ml, respectively; Lanes 8, 9 and 10: controls with BSA at 500, 250, and 125 μg/ml, respectively; Lane 11: control plasmid pRIBOTEX DNA; Lanes 12, 13 and 14: BSA incubated with the probate DNA, but without DTT in the solution (not exposed to hydroxyl radicals); Lanes 1 and 15: reference Hyperladder II