Figure 3

Results of in situ hybridization and immunohistochemistry on thin adjacent section to detect expression of HIF-1α1.2 mRNA and HIF1α protein in malignant and benign prostate tissue. In situ hybridization (antisense probe, Fig. 3a) and immunostaining with HIF1α Ab2 (Fig. 3c) on thin adjacent sections of NE-differentiated prostate adenocarcinoma showed co-localization of HIF1α1.2 transcript and HIF-1α protein. Incubation with sense probe did not generate any detectable hybridization signals (Fig. 3b). Both In situ hybridization (Fig. 3d antisense) and HIF-1α Ab2 immunostaining (Fig. 3f) were negative in non-NE-differentiated prostate adenocarcinoma. In situ hybridization with sense probe performed on non-NE-differentiated prostate cancer (Fig. 3e) was negative. In situ hybridizering on benign prostate tissue showed HIF1α1.2 transcript in NE-like cells of benign prostate tissue (Fig. 3g, ↗). The sense probe on thin adjacent section generated no signals (Fig. 3h,↗). Furthermore, co-localization of HIF1α1.2 transcript (Fig. 3i,↗,◢,↖) and HIF1α protein, detected with HIF1αAb3 (Fig. 3j,↗,◢,↖) was also shown in NE-like cells of benign prostate tissue. Panels a, b, c, d, e, f, g, h, i, j: 40× objective.