Figure 3
Overexpression of HA-Pex11β in A6 cells increased peroxisome numbers. A, D, G are untransfected cells, while B, E, H have been transfected with HA-Pex11β. G and H are additionally transfected with GFP-SKL. Using identical imaging and photography parameters, indirect immunofluorescence using a catalase antibody revealed lower levels of immunofluorescence in untransfected cells (A) versus transfected (B) cells. Similarly, indirect immunofluorescence using a PMP70 antibody revealed lower levels of signal in untransfected cells (D) versus transfected (E). Direct fluorescence for GFP revealed a diffuse signal from GFP in HA-Pex11β untransfected cells, (G) versus the presence of punctate structures in HA-Pex11β transfected cells (H). All images were captured using identical fluorescent settings. The relative fluorescence intensity in 10 regions of twenty randomly imaged cells was quantified using Northern Eclipse software. Graphs on the right represent the average fluorescence intensity of untransfected versus HA-Pex11β transfected cells. Values presented are the means ± SE. Significance at P < 0.05 was determined using Student's t-test, n = 25.