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. 2019 Nov 21;294(52):20196–20206. doi: 10.1074/jbc.RA119.007980

Figure 2.

Figure 2.

Subcellular localization of PP2A Bα and flotillin-1. A, immunofluorescence staining of control BPAECs and BPAECs treated with PMA (1 μm, 30 min), Gö6976 (1 μm, 30 min), OA (5 nm, 30 min), nonspecific siRNA (nonsiRNA), or PP2A Bα–specific siRNA (50 nm) using anti-flotillin-1 (red) and anti-PP2A Bα (green) primary antibodies. Nuclei of cells were stained with DAPI (blue). Scale bars = 25 μm; r values (m–q) are Pearson's cross-correlation coefficients indicating colocalization of flotillin-1 and PP2A Bα. B, magnified region (marked by the white rectangle in A, o) of Gö6976-treated BPAECs. Arrows indicates flotillin-1 in the membrane region. C, cytoplasmic and membrane fractions were isolated from control BPAECs and BPAECs treated with PMA (1 μm, 30 min), Gö6976 (1 μm, 30 min), or OA (5 nm, 30 min). Total lysate and cell fractions were analyzed with anti-flotillin-1, anti-actin (cytoplasmic marker), and anti-CD31 (membrane marker) antibodies. WB, Western blot.