FIGURE 5:

Y14 modulates the formation of processing bodies. (A) HeLa cells were transfected with siRNA targeting to Y14 or eIF4AIII. Indirect immunofluorescence was performed using antibodies against Y14, eIF4AIII, Dcp1a, and Edc4. Arrowhead and arrow, respectively, indicate Y14- and eIF4AIII-depleted cells. Scale bar, 8 μm. Cell lysates were subjected to immunoblotting with antibodies against Y14, eIF4AIII, or α-tubulin. Immunoblotting shows the level of Y14 and eIF4AIII in mock-transfected and siRNA-transfected cells; α-tubulin was used as control. The bar graph shows the average number of P-bodies in mock, Y14-depleted (siY14), and eIF4AIII-depleted (sieIF4AIII) cells (n > 200); asterisks indicate p < 0.01. (B) HeLa cells were transiently transfected with an expression vector encoding FLAG-tagged Y14 (WT), Y14SA, or Y14SE. Immunofluorescence was performed using anti-FLAG and anti-Edc4; arrowhead indicates Y14-overexpressed cells. Immunoblotting was performed using anti-Y14 to detect both FLAG-tagged Y14 (wild-type and mutants) and endogenous Y14 proteins. The bar graph shows the average number of P-bodies in mock and different Y14-overexpressing cells (n > 200).