Immunoblots detect efficient reduction of plectin and complex alterations of integrin expression in A431 clones stably transfected with plectin shRNAs.

<p>Complete cell lysates were obtained from AK13-1-derived subclones that were either stably transfected with scramble shRNA (controls) or plectin shRNA (shPlectin). For comparison, cell lysates from non-transfected A431 cells were prepared. The cells had been cultivated on laminin 332-rich matrices for 48 h either without (left) or with FCS (right). Proteins were separated by 8% SDS-PAGE. Note the efficient down regulation of plectin by 80–90% in both plectin shRNA clones as detected by two different antibodies (*guinea pig, **mouse). In addition, integrins β1 and α3 are also noticeably reduced in plectin-deficient cells. Integrin β4 is only reduced in serum-starved cells upon plectin depletion. The changes in expression levels of other integrins are apparently not linked to plectin expression. Note also, that integrin expression is increased in the presence of serum.