Effects of siRNA knockdown of selected P-body interacting proteins. (A) Immunofluorescence microscopy and Western blot results showing a significant loss of P-bodies in mouse NIH3T3 cells when Pan3 (and not Pan2) was knocked down by specific siRNAs. P-bodies were visualized using rabbit anti-Rck/p54 antibody followed by secondary goat anti-rabbit conjugated to Alexa 488 (green). Cell nuclei were stained by DAPI. (B) Immunofluorescence microscopy results showing effects on P-bodies when the indicated endogenous proteins were knocked down by specific siRNAs in human U2OS cells. P-bodies were visualized with rabbit anti-Rck/p54 antibody followed by secondary goat anti-rabbit conjugated to Alexa 488 (green). Cell nuclei were stained with DAPI. Western blots showing effective knockdown of the targeted endogenous proteins. (C) The changes in P-body number following knockdown of target proteins were analyzed. The quantification of P-bodies was determined by counting the number of P-bodies of a group of cells per experiment. Data are presented as the mean P-body number per cell ± standard error of the mean (n > 4). The asterisk denotes a significant difference analyzed by a paired t-test.
