Fig. 8a, b

Effect of ZC3H41 or Z41AP depletion on protein synthesis. a Immunoblot showing puromycin incorporation into nascent polypeptides. Uninduced cells not treated with puromycin (− puro), uninduced cells incubated in the presence of cycloheximide 30 min prior to treatment with puromycin (CHX), uninduced cells treated with puromycin (− tet), cells treated with puromycin after 48 h of RNA induction (+ tet). Puromycilated polypeptides were detected using an anti-puromycin antibody. DRBD3 was used as a loading control. b Quantification of the puromycin signal from the immunoblot. Percentages of puromycilated polypeptides upon depletion by RNAi relative to those measured in uninduced cells are expressed as the mean (horizontal lines) ± SEM (shaded areas) of three independent RNAi inductions (dots). The horizontal dashed line indicates 100% puromycin signal relative to uninduced cells (i.e. no changes in translation). Two-sided Student’s t-tests (n = 3) were used to assess whether translation rates were significantly altered upon depletion of PuREBP1, ZC3H41 or Z41AP. * P < 0.05; differences observed upon depletion of PuREBP1 were found to be not significant (P = 0.56). For other abbreviations, see Figs. 1 and 2