Figure 4
Target modulation of proliferation and survival proteins by AGR2. (a) Lysates 72 hours after transfection with nontargeting control (iNC) or AGR2 (iAGR2) were evaluated with Western blot for modulation of regulators of growth and survival. Note: Some blots may be from different gels run with the same set of samples. (b) ZR-75-1 cells were treated for 6 hours with BSA (5 μg/mL), Novus rhAGR2 (rhAGR2 (N)), or in-house rhAGR2 (rhAGR2(I)), and analyzed with Western blot for cyclin D1 induction. (c) ZR-75-1 cells were plated in eight-well chamber slides and treated with 5 μg/mL of BSA or rhAGR2 (I) for 6 hours. Cells were stained with cyclin D1, and mounting medium containing DAPI was used. Images were taken by using a fluorescent microscope and pseudo-colored in Adobe Photoshop. (d) Quantitation of cyclin D1 immunofluorescence images. The percentage of cells in each bin based on cyclin D1 intensity is represented (Bin 1, weakest staining; Bin 4, brightest staining). Results are expressed as the mean ± SD; n = 4.