Figure 2.

Compound 147 reduces lysate levels of ALLC in ALMC-2 cells. (A) Graph showing normalized amounts of ALLC in lysates prepared from ALMC-2 cells incubated for 18 hours with vehicle (Veh) or 147 (10 µM). ALLC was quantified by ELISA. Error bars show standard error of the mean (SEM) for 23 replicates across 5 independent experiments. (B) Representative autoradiogram of [³⁵S]-labeled ALLC immunopurified from ALMC-2 cells treated for 18 hours with Veh or 147 (10 µM) and then labeled for 30 minutes with [³⁵S]. The experimental protocol is shown above. (C) ALLC quantification of autoradiograms shown in panel B normalized to Veh-treated cells. Error bars show SEM for 5 independent experiments. (D) Autoradiogram of whole-cell lysates prepared from ALMC-2 cells treated for 18 hours with Veh or 147 and then metabolically labeled with [³⁵S] for 30 minutes. (E) Quantification of ALLC in conditioned media prepared on ALMC-2 cells treated for 18 hours with Veh, 147 (10 µM), and/or ISRIB (200 nM), as indicated. ALLC was measured by ELISA and normalized to Veh-treated cells. Error bars show SEM for 14 replicates across 3 independent experiments. (F) Quantification of ALLC in lysates prepared from ALMC-2 cells treated for 18 hours with Veh, 147 (10 µM), and/or ISRIB (200 nM), as indicated. ALLC was measured by ELISA and normalized to Veh-treated cells. Error bars show SEM for 14 replicates across 3 independent experiments. **P < .01 (paired Student t test); ***P < .005 (unpaired Student t test).