Western blot analysis of cytoplasmic and nuclear IGF1R in MCF10A cells.

<p>MCF10A cells were transfected with IGF1R siRNA (10 nM) or non-targeting (NT) siRNA and harvested 48 hr post transfection. (A) Western blot analysis of total IGF1R, Sumo-1, tubulin [a marker for the cytoplasmic <b>(C)</b> fraction] and SP1 [a marker for the nuclear <b>(N)</b> fraction]. (B) Densitometric analysis of Western blots. The results of a representative experiment repeated three times with similar results are shown. Analyses were done using the “ImageJ software”. Blots of the cytoplasmic fraction were exposed for short periods (2–5 min) whereas blots of the nuclear fraction were exposed for longer periods (10–15 min). Thus, it is not possible to compare between relative abundance of proteins between fractions.