Figure 1.
Mice treated with 25 mg/kg of PQ7 were euthanized at 6, 12, 24, and 36 hours. The total amount of PQ7 administered to each animal was defined as 100%. Bar graph represents the mean distribution of PQ7 with a 95% confidence interval. Data obtained from sample size of n = 6 mice.
Figure 2.
Effect of PQ7 on connexin 43 expression in normal tissue.
A) Immunohistochemisty of tissue sections. Paraffin-embedded sections stained with antibodies against the gap junction protein Cx43in female C57BL/6J organs harvested after a single IP injection of PQ7 (25 mg/kg) at 6, 12, 24, and 36 hours. Proteins staining: brown, counterstaining: blue (hematoxylin). Images represent only 1 of n = 6 per group at a 100X magnification. Scale bar = 10 µm. B) Graphical representation of western blot analysis examining the effect of 6, 12, 24, and 36 hours of PQ7 treatment on the level of Cx43 expression. Mice without PQ treatment were used as a control. Bar graph shows the pixel intensities of protein bands normalized to the pixel intensities of loading control protein (actin) as a percentage of the control tissue. * P-value < 0.05 compared to control.
Figure 3.
Tumor growth (mm3) in PyVT female mice.
Tumors measured in two dimensions with calipers every 2 days prior to administration of treatment for panels A and B) Pre, C and D) Early, and E and F) Late stages of tumor development. Panels A, C, and E) The tumor size is expressed over the 14 day treatment period for the DMSO (control) and PQ7 (25 mg/kg) treated PyVT mice. Days 0-12 represent the days of the 7 IP injections, day 14 represents the end of the study with measurements prior to tissue harvest. Panels B, D, and F) The overall change in tumor size after no treatment, or treatment with DMSO (control) or PQ7 (25 mg/kg) via 7 IPs. * P-value < 0.05 compared to controls.
Figure 4.
Number of developed tumors in PyVT female mice during development.
Tumors identified grossly during the A) Pre, B) Early, and C) Late stages of tumor development after a 14 day period with either no treatment, or treatment with DMSO (control) or PQ7 (25 mg/kg) via 7 IPs. * P-value < 0.05 compared to controls.
Figure 5.
Analysis of tumors isolated from PyVT females 48 hours after the last IP injection.
A) Quantitative analysis of PQ7 in the tumor homogenate. Data obtained from a minimum of three samples per developmental period. Data points represent the nanomolar concentration of PQ7 in each tumor isolated from treated mice, while the dashed lines represent the mean concentration of the PQ7 in all the tumors analyzed. B, C, and D) Graphical representation of protein expression in tumors from Western blot analysis. Fold-pixel intensity of B) Cx43, C) Cx46, and D) PKC-α normalized to loading control in PyVT female tumors treated with DMSO (control) or PQ7 (25 mg/kg) via 7 IPs in each of the three stages of tumor development. n = 4. * P-value < 0.05 compared to control.
Figure 6.
Immunohistochemisty of tumors from PyVT females.
Paraffin-embedded tumor sections stained with A) H&E or antibodies against B) Cx43, C) Cx46, and D) PKCα from PyVT females treated with DMSO (control) or PQ7 (25 mg/kg) via 7 IPs at either Pre, Early, or Late stage of tumor development. Proteins staining: brown, counterstaining: blue (hematoxylin). Images represent only 1 of n = 6 per group at a 100X magnification. Scale bar = 10 µm.