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The “ATF4 5: 5′ATF4.GFP” plasmid (#21852) was purchased from Addgene (52). MEFs were transfected with the ATF4-GFP–expressing plasmid using jetPRIME transfection reagents (Polyplus-transfection) and, 24 hours later, were pretreated with 5 μM ONC201 for 24 hours and 1 μM thapsigargin for 1 and 6 hours. DAPI-negative and GFP-positive cells were measured by flow cytometry.
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