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. 2006 Mar 30;97(4):259–270. doi: 10.1111/j.1349-7006.2006.00169.x

Figure 5.

Figure 5

Alteration of the expression of filamentous‐actin (F‐actin) and actin binding proteins (ABP) by constitutive expression of the WT1 17AA(–)/KTS(–) isoform. (a) WT1 17AA(–)/KTS(–) isoform‐transduced or control vector‐transduced TYK cells were stained immunocytochemically for F‐actin (red), the focal adhesion protein vinculin (green) and the nucleus (blue). (b) Cell lysates of control vector‐transduced (lanes 1 and 2) or WT1 17AA(–)/KTS(–) isoform‐transduced (lanes 3 and 4) independent cell clones were immunoblotted with antibodies specific for total actin, F‐actin, α‐tubulin or glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH; used as a loading control). (c) Expression of mRNA of ABP in control vector‐transduced (n = 3, lanes 1–3) or WT1 17AA(–)/KTS(–) isoform‐transduced (n = 3, lanes 4–6) TYK cell clones isolated independently was analyzed by reverse transcription–polymerase chain reaction under the conditions shown in Table 1. (d) Cell lysates of control vector‐transduced (lanes 1 and 2) or WT1 17AA(–)/KTS(–) isoform‐transduced (lanes 3 and 4) TYK cell clones were immunoblotted with antibodies specific for α‐actinin 1, cofilin and gelsolin. (e) WT1 17AA(–)/KTS(–) isoform‐transduced or control vector‐transduced TYK cells were stained immunocytochemically for α‐actinin 1, cofilin and gelsolin (green). The nucleus of cells was stained with propidium iodide (red). (a,e) Scale bars = 10 µm. (f,g) Cell lysates of control vector‐transduced or WT1 17AA(–)/KTS(–) isoform‐transduced HT‐1080 (f) or TE10 (g) cell clones were immunoblotted with antibodies specific for α‐actinin 1, cofilin and gelsolin.