Classification of RAD51-focus morphology upon hyperthermia in human bladder cancer.

A) Two hours after DNA-damage induction irradiation or zeocin, tumors were fixed and prepared for immunohistochemistry and subsequently stained for GMNN (green) and RAD51 (red). Pictures presented are examples of both undissociated and dissociated tumors exhibiting the two distinct morphological responses of RAD51-foci. Panels below the blue thermometer received no treatment before damage induction; panels below red thermometers were first treated with hyperthermia. The images are maximum projections of Z-stacks from representative areas of the tumors. B) Graph representing the frequency distribution of the morphological response of RAD51-focus upon hyperthermia treatment. The white part of the bar represents foci which have become smaller and more dispersed upon hyperthermia, the black part represents foci that did not change in morphology. Each bar contains the number (n) of analyzed tumors below. C) Frequency distribution based on the tumor preparation step (undissociated or dissociated) or on the method of damage induction. Two tumors are classified in both methods of tumor preparation, because both were performed on the same tumor, and had the same result. No differences were found in the foci-response distribution between the technical methods (Fisher’s Exact test, p-values stated in the Fig). D) Distribution of RAD51-foci response upon hyperthermia based on clinical data (available for 36 tumors). No differences were found in the frequency distribution based on tumor grade (G1 and G2 vs G3, Fisher’s exact test), tumor stage (Ta, T1 and T2, Chi-square), primary versus recurrent tumors (Fisher’s exact test), or in the EAU risk classification (intermediate or high [43], Fisher’s exact test). p-values are indicated in the figures.