Figure 5

Limited differential expression in Rab5-overexpressing PCF cells. Panel A: Western blot analysis for the clathrin heavy chain (CLH) and Rab5A, for wildtype BSFs and PCFs, PCFs overexpressing Rab5A (5AWT), Rab5A^QL (GTP-locked mutant) and Rab5B^QL (GTP-locked mutant) cell lines. BiP was used as a loading control. Panel B: Scatter plots of raw data for representative microarrays for BSF (Cy5) vs PCF (Cy3) experiment (plot as in Figure 1 for comparison), and Rab5-overexpessing lines (Cy5) vs PCF (Cy3) as indicated. Cy5 fluorescence is plotted on the Y-axis and Cy3 fluorescence on the X-axis. Spots with a Cy5/Cy3 or Cy3/Cy5 ratio above two are highlighted in black, spots with a ratio below two are shown in grey. Panel C: Relative expression levels of Rab5A, Rab5B, CLH, RabX3, COPIε, and adenylate kinase 3 (AK3) in wildtype SMBs and PCFs, and in the Rab5-overexpressor PCF lines, as assessed by qRT-PCR.