Figure 7

Figure 7

ROS generation and alkalization of intracellular pH induced by CRPs. (A) Effect of CRPs to generate oxidative stress in vitro. ROS formation was quantified in ZF4 cells transfected with pMCV1.4-crp1-7 for 48 h and incubated for 30 min with the stress indicator CM-H2DCFDA. ROS generation was determined measuring fluorescence intensity (n = 4). (B) Ability of the CRP-mix to modulate the pH of lysosomes. Changes in the lysosomal pH were determined in ZF4 cells co-transfected with each crp-encoding plasmid for 48 h and stained with LysoTracker Green DND-26 for 30 min. The quantification of the green fluorescence was carried out by flow-cytometry (n = 6). Results are shown in arbitrary units (a.u.). All statistically significant level differences between treatment and corresponding control groups are indicated with symbols as in Fig. 1. Data were analysed by using two-tailed unpaired Student’s t-test.