Analysis of nmMHC redox state and β-actin association in human fibroblasts. Cells lysates from detached and spread cells, treated with or without NDGA, were labelled with BIAM and nmMHC was immunoprecipitated with specific antibodies. (a) BIAM labelled pattern of nmMHC was revealed by western blot using HRP-streptavidin conjugate; amount of immunoprecipitated nmMHC was obtained probing the membrane with anti-nmMHC antibodies; the histogram shows the ratio between two corresponding samples after densitometric evaluation. versus rounded. (b) Representative immunoblot showing actin glutathionylation in rounded and spread human fibroblasts. The histogram corresponds to the ratio between GSH-actin and total actin. ^♦ versus rounded. (c) Analysis of nmMHC-actin association. An anti-nmMHC immunoprecipitation was performed from rounded and spread cells treated with or without NDGA. The detection of actin associated with nmMHC was obtained with antiactin immunoblot, while anti-nmMHC immunoblot was used for normalization. The histogram corresponds to the ratio between the values obtained by densitometric analysis of two corresponding samples of the blots. versus rounded. Similar results were obtained in four independent experiments. a.u.: arbitrary units.