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Fig. 7. P-cortactin anchors actin stress fibers in COS-7 and NIH3T3 cells at interphase. (A) NIH3T3 cells were stained for P-Tyr421-cortactin (PY⁴²¹C), cortactin (Cort) or F-actin (using phalloidin); nuclei were stained with DAPI. M, merged images. Scale bars: 50 µm. (B) NIH3T3 cells were stained for P-Tyr421-cortactin (PY⁴²¹C), P-Tyr466-cortactin (PY⁴⁶⁶C) or P-Tyr482-cortactin (PY⁴⁸²C) together with anti-paxillin (PAX) and F-actin (using phalloidin); nuclei were stained with DAPI. M, merged images. Scale bars, 50 µm. (C) Merged images of HIN3T3 cells stained for F-actin together with cortactin or P-cortactin. Scale bars, 10 µm. (D) COS-7 cells stained for P-Tyr421-cortactin (PY⁴²¹C), paxillin (PAX), or F-actin (using phalloidin); nuclei were stained with DAPI. M, merged images. Arrowheads indicate centrosomes associated with P-cortactin. Scale bars, 50 µm (in interphase cells, top panels), 10 µm (in metaphase cells, bottom panels). (E) Co-immunoprecipitation of P-cortactin and actin. 3T3-L1 cells were homogenized and sonicated. The cell lysate was immunoprecipitated (IP) using anti-cortactin antibody (-Cort) or anti-phosphotyrosine⁴²¹ cortactin antibody (-PY⁴²¹C). IB, western blot; T, total cell lysate; IP, immunoprecipitated sample.

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