β‐actin depletion from mouse embryonic fibroblasts results in an altered transcriptional response rendering these cells a myofibroblast like phenotype. The proteins and upstream regulatory factors responsible for this acquired phenotype, with prominent focal adhesions and stress fibres, are unknown. Data‐mining of the changed proteome revealed that actin binding proteins associated with stress fiber or focal adhesion formation are overexpressed in the β‐actin knock‐out cells and that many of these contain CH‐, LIM‐ or EFh‐ domains. Furthermore in silico analysis predicts potential common upstream regulators that may, at least partly, coordinate the altered transcriptional response. © 2013 Wiley Periodicals, Inc.