Figure 3. Aberrant HIF1α expression in PDAC cancer associated fibroblasts.
A. The level of MCT4 was determined in the indicated established pancreatic cancer cell lines and CAF cultures by immunoblotting. B. The level of MCT1 was determined in the indicated established pancreatic cancer cell lines and CAF cultures by immunoblotting. C. HIFα and MCT4 levels were determined by immunoblotting following the knockdown of HIF1α with RNAi or the induction of hypoxia through the use of CoCl2. D. The stability of HIF1α was evaluated in CAFs following treatment with cycloheximide. E. HIF1α was detected by immunoblotting in the presence of cycloheximide in the absence or presence of the proteasome inhibitor MG132. F. HIF1α protein levels were determined in the absence and presence of DMOG. G. RNA levels of the indicated genes as determined by microarray analyses (**p < 0.01 for tumor cell line vs. CAF). H. MG132 was used to interrogate the synthetic rate of HIF1α protein in the indicated cell lines. I. The expression of HIF1α target genes was evaluated in the indicated cell lines by microarray analysis (**p < 0.01 for tumor cell vs. CAF).