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Fig. 2. Immunoblot of larger palladin isoforms. (A) Tissues were dissected from a neonatal mouse and immunoblotted with the polyclonal antibody a-4IgNT that detects the N-terminal extension of the 200 kDa and 140 kDa palladin isoforms. This antibody does not crossreact with the 90 kDa isoform. Similar to the reported expression of the 90 kDa isoform, the 140 kDa isoform of palladin is expressed in most neonatal mouse tissues, whereas the 200 kDa isoform is predominantly restricted to heart, muscle and bone. (B) The 140 kDa isoform is considerately downregulated in many adult tissues, but is abundant in tissues rich in smooth muscle, such as stomach and uterus. The 200 kDa isoform is expressed in striated muscle of the adult mouse. (C) Vector-driven expression of the putative 140 kDa palladin nucleotide sequence in COS-7 cells results in the translation of an immunoreactive band with the same apparent molecular mass observed for endogenous protein in astrocytes. Astrocytes express relatively high levels of the 140 kDa isoform of palladin. Immunodetection with a previously characterized monoclonal (left blot) that reacts with the 90 kDa, 140 kDa and 200 kDa isoforms demonstrates that the 90 kDa and 140 kDa isoforms are expressed in cultured astrocytes. The polyclonal antibody a-4IgNT (right blot) specifically detects the 140 kDa isoform in these cells.

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