Fig. 3. Quantitation of LPA₁ internalization. LPA₁-transfected cells were incubated with 10 µM LPA for various times and then fixed and processed for quantitation of receptor internalization by laser scanning confocal microscopy as described in the Materials and Methods. (A) A representative confocal image is shown of untreated and LPA-treated cells stained with ConA to label the PM, and anti-FLAG antibodies to label FLAG-tagged LPA₁. Note that LPA₁ extensively co-localizes with ConA in untreated cells, but localizes to punctate fluorescent structures after LPA treatment. Bar, 10 µm. (B) Quantitation of internalization showed that approximately 40% of LPA₁ is internalized within 15 minutes after LPA treatment. The data is presented as the mean ± s.e.m. at each time point (n=24 cells analyzed). ***P<0.0001 compared with untreated cells.

Return to article