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Figure 5 Reverse transcriptase-polymerase chain reaction (A) and Western blots (B) of procaspase-1, -3, -7, -8, -9, -12, XIAP, and Smac/DIABLO RNA and protein levels in cardiac tissues from hearts, exposed and unexposed, to ischemic/reperfusion (I/R), with and without in vivo treatment with minocycline. Corresponding densitometric analyses are shown at the bottom of each panel. Values are represented as relative fold change of the control value set to 1. Statistical analysis (*p < 0.05; **p < 0.01; ***p < 0.001) was performed versus control hearts (C) either unexposed (non-ischemic hearts treated in vivo with minocycline [Mnc]) or exposed to I/R (ischemic hearts pretreated in vivo with minocycline). (C) Caspase-3, -7, -8, and -9 enzymatic activity in tissue extracts from non-ischemic and ischemic control hearts and hearts pretreated in vivo with minocycline and exposed to I/R. Data are expressed as mean ± SD. ***p < 0.001 vs. control hearts. (D) Expression levels of cytochrome c and Smac/DIABLO proteins in mitochondrial and cytosolic fractions from control hearts exposed and unexposed to I/R, with and without in vivo treatment with minocycline. HSP60 and actin were used as internal controls for the mitochondrial and cytosolic fractions, respectively. The correspondent densitometric evaluation is shown at the bottom of the panel. Values are represented as relative fold change of the control value set to 1. Statistical analysis was performed as stated above. The reverse transcription-polymerase chain reaction and Western data are representative of five experiments performed in individual animals, and the in vivo treatment was performed as described in the Methods section and the legend to Figure 3.