Figure 4: Antagonizing the Function of CTGF mAb During Cardiac I/R Injury (Study III)

Mice were treated with IgG vehicle or CTGF mAb and subjected to ischemia−reperfusion injury (I/R). (A) Quantitative analysis of TUNEL-positive cells in hearts subjected to 30 min of ischemia and 3 h of reperfusion is shown. TUNEL-positive cells are marked with arrows; scale bar: 20 μm. (B) Mice were treated with IgG vehicle or CTGF mAb 24 h before ischemia and at reperfusion, or with CTGF mAb at reperfusion only. Infarct size and area at risk were analyzed from triphenyl tetrazolium chloride (TTC)−stained myocardial sections. (C) Western blot analysis of samples from infarct area 3 h after I/R injury. Analysis of phosphorylated protein kinase B (Akt), ERK, JNK, signal transducer and activator of transcription 3 (STAT3), p38, protein kinase C alpha (PKCα), SMAD2, and SMAD1/5 is shown. GAPDH was used as a loading control. Ratio of p-JNK to GAPDH and p-STAT3 to GAPDH data in the bar graphs are presented as mean ± SD. Number of animals in 3-h I/R experiment, including TUNEL labeling, was sham (n = 3), IgG (n = 6), and CTGF mAb (n = 6). Number of animals in 24-h I/R experiment including TTC staining was IgG (n = 11), CTGF mAb (n = 12), and CTGF in reperfusion (n = 12). TUNEL = terminal deoxynucleotidyl transferase dUTP nick end labeling; other abbreviations as in Figures 1 and 2.