<b>TIA1 promotes ULK1 protein expression and autophagy during nutrient starvation</b>

(A) Loss of TIA reduces ULK1 protein expression and autophagy: Calu-1 cells transfected with either siRNA (control) or siRNA specific to TIA1 were subjected to serum starvation for the indicated times with or without treatment with bafilomycin A1. Cells were harvested after the indicated times and protein levels of ULK1, TIA1 and LC3 were determined by immunoblotting. ACTB was used as a loading control. (B) Quantification of ULK1 protein levels in A. Data are representative of three independent biological replicates, showing mean +/- SD. **** p value &lt;0.001. (C) TIA1 knockdown does not affect ULK1 transcription: Total RNA was extracted from cells harvested under the conditions indicated in a and ULK1 mRNA abundance was determined by qRT-PCR. GAPDH was used as a control. Data are representative of three independent biological replicates. (D) Quantification of LC3-II levels in a. Data are representative of three independent biological replicates, showing mean +/- SD. Two-way ANOVA was used to determine statistical significance. **** p value &lt; 0.01, *** p value &lt;0.05. (E) TIA1 binds to the 3' UTR of ULK1: RNA immunoprecipitation was performed using TIA1-specific antibody and the mRNA bound to the protein was quantified by qRT-PCR using two primers spanning different regions in the 5' UTR and 3' UTR. Data are representative of three independent biological replicates, showing mean +/- SD. One-way ANOVA was performed to determine statistical significance. ** p value = 0.0013, * p value = 0.0358.