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. 2010 Apr 15;21(8):1409–1422. doi: 10.1091/mbc.E09-06-0451

Figure 4.

Figure 4.

Knockdown of h3/acidic CaP does not affect PKCα targeting upon PDBu stimulation. (A) REF52.2 cells were transfected with siRNA against aCaP siRNA (i–p) or nontargeting control siRNA (a–h). Three days after transfection, cells were treated with 0.01% DMSO for control (a–d and i–l) or with 1 μM PDBu for 30 min (e–f and m–p) before staining for aCaP (a, e, i, and m) and PKCα (b, f, j, and n) with the rabbit polyclonal anti-h3/acidic CaP and the mouse monoclonal anti-PKCα antibodies. Images were obtained with deconvolution microscopy. The yellowish signal in the merged images indicates colocalization (d, h, l, and p). Bar, 10 μm. (B) REF52.2 cells transfected with siRNA against h3/acidic CaP or with nontargeting control siRNA were treated with 1 μM PDBu for 30 min. After an immunofluorescence staining for h3/acidic CaP and PKCα, cells showing a PKCα translocation to either the cell cortex or podosome-like structures were counted. The graph represents three independent experiments, where at least 100 cells were counted.