Figure 8: Assessment of nuclear translocation of FoxO3a and p53 redox-response element in MSCs challenged with LPS. Counterstaining of nuclear DNA was with Hoechst 33342 (blue channel). FoxO3a staining is in red (control (a1) and (a2) versus LPS treatment (b1) and (b2)). ((a2)-(b2)) overlay of confocal projections of FoxO3a protein (red channel) and nuclear DNA (blue channel). FoxO3a nuclear translocation is indicated with arrows in (b1) and (b2), where nuclear fraction of FoxO3a appears in pink color due to interference of “red” and “blue”. ((c1), (c2), (d1), (d2)) Confocal projections of p53 protein (red channel) in MSCs (control (c1) and (c2) versus LPS (d1) and (d2)). Counterstainings were done with Hoechst 33342 (for nuclear DNA, blue channel) and anti-TOM20 IgG (for mitochondria, green cannel). (c2) and (d2) are overlay of confocal projections of p53 protein (red channel), TOM20 protein (green channel), and nuclear DNA (blue channel). Note that there was no detectable increase in the p53-immunofluorescence in nuclear areas of LPS-challenged MSCs. The confocal images were taken with pinhole setup to obtain 0.5 μm Z-sections. Experimental conditions were the same as indicated in Figure 2.