Fig 2: Microphotographs of coronal sections through the rostral (A,B) and middle (C–F) regions of the dentate gyrus of control (A,C,E) and tbl mice (B,D,F) to illustrate the immunoreactivity to caspase 3 and the neuronal marker proteins HuC/HuD. Almost all caspase-3 immunoreactive cells are located within the hilus, the subgranular zone and the granule cell layer, while HuC/HuD also labels interneurons located in the molecular layer. In the control DG it is possible to find caspase-3 labeled cells that do not co-express HuC/HuD (A,C, arrows), whereas in the tbl DG, caspase-3 immunoreactive cells also are labeled by the HuC/HuD antibody (B,D, arrows). Quantitative analysis demonstrates that the tbl DG has significantly more caspase-3-HuC/HuD (G, ***p = 1.902245E–12) and cleaved caspase-3-HuC/HuD double labeled neurons than the controls (H, ***p = 5.524E–06). gcl, granule cells layer; h, hilus; mol, molecular layer; sgz, subgranular zone. Bar = 200 µm (A–F), and 30 and 60 µm (F).

Fig 3: Dysfunction of non-stimulated and LPS-stimulated HIV neutrophils. (A) Expression of CD11b and CD18 adhesion molecules on the surface of neutrophils in HIV vs. HCs. The bars represent the mean of fluorescence intensity ± SD (B) Reactive oxygen species production by non- and LPS-stimulated neutrophils. The bars represent the mean of ROS production in Relative Fold Units ± SD. (C) The comparison of the profile of cytokine/chemokine/growth factors released by non-stimulated (left panel) and LPS-stimulated (right panel) neutrophils. The blue line determines the border between up- and down-regulated factors. (D) ICC double labeling for caspase-3 (red pseudocolor) and annexin-V (green) revealed that circulating HIV neutrophils are apoptotic. Additional LPS stimulation does not inhibit neutrophil apoptosis as it takes place in HC neutrophils. Nonspecific fluorescence (signal noise) was electronically diminished to the level when nonspecific signal was undetectable (background). The bars represent average fluorescence intensity ± SD calculated from four patients, using at least 100 single cells for each test.

Fig 4: DEN induces AML cell apoptosis. (A) AML cells (n = 10) were treated for 48 h with increasing doses of DEN. Annexin V/PI staining was used to detect apoptosis. (B) Representative FACS analysis of apoptosis after treatment with 1 mM DEN. (C) FACS analysis of active caspase-3 expression in AML cells after treatment with 1 mM DEN for 48 h (n = 6). (D) Representative overlay histograms of Active caspase-3 expression. (E) Immunofluorescence analysis of activated caspase-3 (green) after 48 h of treatment with 1 mM DEN. Nuclei were counterstained with DAPI (blue). 40X magnification, scale bar 20 µm. (F) FACS analysis of mitochondrial membrane potential of AML cells after 48 h of exposure to 1 mM DEN. The histogram shows the percentage of JC-1 aggregates (cells emitting red fluorescence in the FL-2 channel) and JC-1 monomers (cells emitting green JC-1 detected in the FL-1 channel) (n = 6). (G) Representative dot plots of JC-1 staining. Data are expressed as mean ± SD. Statistical analysis was performed by using ANOVA followed by Dunnett's multiple comparison test with untreated group as control and by Student's t-test. *p < 0.05; **p < 0.01.

Fig 5: Photolyase significantly inhibited UVB-induced oxidative stress and inflammatory responses in mice. (A) Lipid peroxidation in the dorsal skin in each group after rPHO treatment was measured by quantifying malondialdehyde (MDA). (B) SOD activity in the dorsal skin after rPHO treatment. (C) GSH-Px activity in the dorsal skin after rPHO treatment. (D–F) Expression of IL-6, IL-1ß, and TNF-a in skin was measured. mRNA relative levels were normalized to Gapdh levels. (G) Expression of caspase 3, Bcl-2, and Bax protein examined by western blotting. (H) Semi-quantification analysis of (G) by ImageJ. Data are expressed as means ± SD (n = 5). * p < 0.05, ** p < 0.01, *** p < 0.001.