Fig. 4

The ribosome stall sensor ZNF598 critically regulates the translational quality control and toxicity of APP.C99. a Immunoblots showing co-IP between APP.C99 and ZNF598 and other ribosome-associated QC factors. b Translocon pull-down with ConA beads showing increased ZNF598 and Ltn1 association with Sec61 translocon in APP.C99 transfected cells. c Immunoblots showing the effect of ZNF598 RNAi or OE on the level of stalled APP.C99. d Immunoblots showing the effect of ZNF598 RNAi on the level of mOC78-positive aberrant APP.C99 species. Scale bar, 5 µm. e MTT viability assay showing the effect of ZNF598 RNAi on APP.C99 toxicity. f Immunoblotd showing the effect of ZNF598 RNAi on the level of stalled APP.C99 in Mhc > APP.C99 fly muscle. g Immunostaining and data quantification showing effect of ZNF598 RNAi on the level of Ub- and 6E10- positive aggregates in Mhc > APP.C99 fly muscle (n = 10). Scale bar, 10 µm. h Effect of ZNF598 RNAi on locomotor activity in elav > APP.C99 flies (n = 20–25). i Effect of ZNF598 RNAi on the lifespan of Mhc > APP.C99 flies (n = 80–100). j Effect of ZNF598 RNAi on the aversive taste memory response in elav > APP.C99 flies (n = 60). k-m Immunostaining (k, l) and data quantification (m) showing effects of ZNF598 RNAi on the level of 6E10-positive aggregates in the muscle of Mhc > APP; BACE1 flies (k, n = 10), or effect of ZNF598 RNAi on the level of 6E10 and Ub double-positive aggregates in the brain of elav > APP; BACE1 flies (l, n = 20–25). Scale bars: 10 µm (k), 20 µm (l). n Effect of ZNF598 RNAi on the aversive taste memory response in elav > APP; BACE1 flies (n = 60–80). Error bars, ± SEM; ^∗P < 0.05, ** P < 0.01. Immunoblots represent at least 3 independent repeats