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Anti-MUC5AC was incubated for 60 min, anti-MUC5B for 30 min and anti-E-cadherin for 30 min

Javadiyan et al

To determine whether ATF4 activation is both necessary and sufficient for mTORC1-mediated regulation of these gene targets related to amino acid acquisition and utilization, we knocked out Atf4 using CRISPR/Cas9 in Tsc2 -/- MEFs and confirmed biallelic disruption (Figure 4A)