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. 2022 May 2;9:809985. doi: 10.3389/fmolb.2022.809985

FIGURE 2.

FIGURE 2

UPR and UPS deregulation induced by recombinant tRNASer(AGA). (A) Expression of tRNASer(AGA) increases BiP expression in BEAS-2B cells. (B) Levels of eIF2α-P were decreased in cells with tRNASer(AGA). (C)tRNASer(AGA) expression in BEAS-2B cell line increased the activation of ATF4 transcription factor. (D) Expression of GADD34 was increased in BEAS-2B cells expressing the recombinant tRNASer(AGA). (E) Immunoblots using antibodies against BiP, eIF2α-P, eIF2α, ATF4-P, ATF4 and GADD34. Β-tub served as protein loading control. Graphics depict average ±SEM (n = 3), with three technical replicates. (F) No differences are observed between BEAS-2B Mock and tRNASer(AGA) cells regarding ubiquitin levels. Graphic represents average ±SEM of five independent experiments. (G) Upregulation of tRNASer(AGA) in BEAS-2B increases proteasome activity by 1.32-fold. Graphic depicts average ±SEM (n = 4), with three technical replicates. (H) Expression of ubiquitin. ß-tubulin represents the internal control. Total protein was extracted from BEAS-2B Mock cells (lanes 1, 2 and 3) and BEAS-2B tRNASer(AGA) (lanes 4, 5 and 6). Image represents results from one independent experiment with all technical replicates. Data was analyzed with One-way ANOVA and Holm Sidak’s post-test and significant p values are shown (*p < 0.05; **p < 0.01; ***p < 0.001).