Figure EV4. Overexpression of ERdj3^QEVV exacerbates Tg‐induced alterations in secretory proteostasis regulation for the destabilized, aggregation‐prone ^FTTTR^A25T (related to Fig 6).

1. Quantification of total ^FTTTR^A25T in conditioned media measured by SDS–PAGE/immunoblotting. The levels of ^FTTTR^A25T in conditioned media are shown relative to vehicle‐treated cells overexpressing ERdj3^WT. Error bars show SEM for n = 5. ***P < 0.005 from a paired t‐test. A representative immunoblot for this quantification is shown in Fig 6B.
2. Representative autoradiogram and quantification of HEK293T [³⁵S]‐labeled ^FTTTR^A25T in conditioned media prepared on HEK293 cells transfected with ^FTTTR^A25T and ERdj3^WT or ERdj3^QEVV, as indicated. The [³⁵S] metabolic labeling protocol is shown above. Error bars show SEM for n = 4. *P < 0.05 from a paired t‐test.
3. Quantification of normalized ^FTTTR^A25T aggregates in conditioned media measured by CN–PAGE/immunoblotting. Normalized ^FTTTR^A25T aggregates were calculated by the following formula: normalized ^FTTTR^A25T aggregates for a given condition X = [aggregate ^FTTTR^A25T condition X/aggregate ^FTTTR^A25T veh‐treated mock‐transfected cells]/[total ^FTTTR^A25T condition X/total ^FTTTR^A25T in veh‐treated mock‐transfected cells]. The data are shown relative vehicle‐treated cells transfected with ERdj3^WT. Error bars show SEM for n = 5. *P < 0.05 from a paired t‐test.