Fig 2: 5-FU resistant pancreatic cancer cells display increased uptake and accumulation of TPCS2a, and show increased ROS production following PCT. TPCS2a accumulation with a increasing TPCS2a concentration and b increasing incubation time. The graphs show the median TPCS2a fluorescence intensities in live cells and show the average of three and two individual experiments, respectively (Error bars = S.E.). c Representative epi-fluorescence microscopy images of live 5-FU sensitive and 5-FU resistant cells harvested 1, 2 and 4 h after TPCS2a incubation. Co-stain of TPCS2a and Alexa488-Dextran is indicated by yellow fluorescence in the rightmost images. The experiment was performed once but was also verified for the B1Q and the B1L cells (data not shown), confirming the data in (a) and (b). The scale bar is 20 μm. d Relative ROS generation measured 1 h after light exposure, with increasing PCT (TPCS2a + light) illumination doses (30–120 s). The graph shows normalized median DCF fluorescence intensities representative of three individual experiments. e RNA levels (relative quantity) of SOD1 and SOD2 in all cell lines, as measured by RT-qPCR. Error bars represent standard deviation. Statistically significant difference between 5-FU sensitive and 5-FU resistant lines (P < 0.05) is indicated by *. f Western blot analysis showing SOD1 and SOD2 protein expression in untreated cells. The blot is representative from at least two individual experiments

Fig 3: Immunoblot analysis of β-Adrenergic Receptor and markers of mitochondrial β-oxidation in the hearts of CoV2 infected, T. cruzi infected, and coinfected huACE2 mice. Western blot images (A) of β-Adrenergic Receptor (β-AR), cytochrome C (Cyto C), and superoxide dismutase (SOD). β-Actin was used as loading control. Fold changes in the protein levels of β-AR (B), Cyto C (C) and SOD (D) were normalized to β-Actin expression and are shown as bar graphs (A dot plot displaying individual data points is presented in Supplementary Figure 4). The error bars represent standard error of the mean. *p < 0.05, **p < 0.01, ***p < 0.001 and ****p < 0.0001 compared to uninfected sex matched mice (n = 4/sex/group) (#p ≤ 0.05 and ###p ≤ 0.001 for comparisons between CoV2 infected and coinfected mice) (M, male; F, female).

Fig 4: Superoxide dismutase activity. Quantitative analyses from young and elderly myoblasts and myotubes for the activity of the superoxide dismutase enzyme, SOD1. The SOD units (U SOD) were calculated considering that 1 SOD unit is defined as the quantity that inhibits the rate of cytochrome c reduction by 50% per ng of protein. The activity of the superoxide dismutase was expressed as U SOD/ng protein. ∗ p ≤ 0.05.

Fig 5: Platelets from 24 month mice are less active, and richer in antioxidant enzymes as compared to platelets from 6- and 12-month old mice. a-b. Levels of sP-selectin and sCD40L in the mice plasma (6 m n = 12, 12 m n = 15, 24 m n = 15). c. Representative platelet aggregation tracings in response to ADP. d. Quantitation of aggregation shown taking 20 Ω (maximal impedance) as the 100% response (n = 4 mice per group). e-f. Representative zebra plots and quantitation showing the percentage of apoptotic cells as measured by the dye TMRE in the platelets. g-h. Representative immunoblot and respective densitometric analysis showing expression of apoptotic proteins in the platelets. i. Intracellular ROS levels measured in freshly isolated platelets (6 m n = 10, 12 m n = 14, 24 m n = 14). j-k. Activity of the enzymatic antioxidants, catalase and SOD (6 m n = 10, 12 m n = 14, 24 m n = 14). l-m. Representative western blotting images and densitometric quantification for the expression of the key antioxidant proteins. Expression is normalized to GAPDH and expressed as fold change over 6 month mice. Experiments unless otherwise specified, performed in a minimum of n = 10 samples per group. Values are expressed as Mean ± SEM. *p < .05; **p < .005; ***p < .001 *p < .05; **p < .005; ***p < .001 determined by one-way ANOVA with Bonferroni post hoc analysis.