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Figure 3


Fig. 3. Co-localization of endogenous proteins during cell spreading. After 30 minutes of spreading on 10 µg/ml fibronectin, A549 cells were stained with antibodies against MHC IIA (a), tubulin (c), MHC IIB (d), vinculin (f), mono-phosphorylated serine-19 in MLC (pS19-MLC; g,j,m), di-phosphorylated threonine-18, serine-19 in MLC (pT18pS19-MLC; h), or supervillin (k,n) and with Alexa-Fluor-350-conjugated phalloidin for visualizing F-actin (b,e,i,l,o). Alexa-Fluor-488-conjugated (a,d,h,k) and Alexa-Fluor-594-conjugated (c,f,g,j) secondary antibodies were used. (m-o) Enlargements of the boxed areas in j-l. (p-r) Cross-sections of signal intensities along the vertical lines in m-o, respectively. Bars, 10 µm.