Fig. 1. siRNA-mediated depletion of lamin A/C, p150^Glued, EB1 or CLIP-170. (A) Cells depleted of lamin A/C, p150^Glued, EB1 or CLIP-170 were probed with antibodies specific for lamin A/C, p150^Glued, EB1 or CLIP-170 as indicated. (B) RNAi-mediated depletion of EB1 is seen with siRNAs designed to target EB1 but also with CLIP-170b designed to target CLIP-170. This was also seen when we resynthesized this siRNA to eliminate cross-contamination as a possible explanation (CLIP-170b2). EB1 is not depleted by other CLIP-170 targeted siRNA duplexes (CLIP-170a, c or d). Notice that one lane of the lower panel has been removed to align the top and bottom panels with one another (shown by the black vertical line). (C) Use of non-fluorescent (top panels) or fluorescent (lower panels) siRNA duplexes targeting lamin A/C shows the efficiency of transfection with the modified calcium phosphate protocol. Both fluorescence images were acquired with the same microscope settings; fluorescence is only seen when fluorescent siRNA duplexes were used (bottom right) and does not result from cellular autofluorescence. (D) Immunofluorescence localization of EB1 in cells depleted of lamin A/C (top) or EB1 (bottom) confirms efficiency of target depletion. Bars, 10 µm.