Fig. 2

a A short list of representative genes relevant to UPR/ER stress-related cellular response identified through enrichment analysis of mRNA-Seq data obtained from UOK262/UOK262WT cell lines incubated with either Asn, Gln, or both for 96 h. b UPR/ER stress response related mRNA-Seq gene expression verified by means of RT-PCR. The first two panels of bars at each diagram represent the amino acids treatment resulted in expression fold change compared to untreated control in UOK262 and UOK262WT respectively. The dotted bar on the right side of each diagram represents the fold change of gene expression in UOK262 vs. UOK262WT cells treated with both amino acids (Asn + Gln). c Confirmative Western blots of UPR/ER stress-related proteins. The UOK262/UOK262WT cells treated with either Asn, Gln, or both for 96 h before cell homogenization. d Western blot analysis of BiP and GRP94 expression and phosphorylation pattern of pJunB proteins in reverse experiment. The UOK262/UOK262WT cells treated with Asn and Gln for 72 h prior to removal either Asn, Gln, or both amino acids for the next 24 h following by cell homogenization. e Western blot of a spliced (active) variant of an XBP1 transcription factor in UOK262/UOK262WT after 96 h of incubation with Asn, Gln, or both. Diagram represents quantitative densitometry of the sXBP1 protein under different conditions. For all Western blot experiments 20 μg of total protein loaded at each well, unless stated otherwise. Actin used as a loading control. Statistical analysis was performed to compare the untreated versus treated samples using one-way ANOVA test following by unpaired, two-tailed t tests (GraphPad Prism v. 8). (**) p < 0.01. The significance between treatment groups shown as (#) p < 0.05