Extended Data Fig. 5: LARP4 is regulated by NR4A1/2 and modulate mRNA translation in exhausted CD8+ T cells.
From: LARP4-mediated hypertranslation drives T cell dysfunction in tumors
(a) Immunoblotting is shown to validate changes in LARP4 expression in control and Larp4-KO TEX cells after 8 days of in vitro chronic antigenic stimulation. (b) Immunoblotting showing the change of LARP4 expression in control and Nr4a1/2-KO Cas9+ T cells after 6 days of in vitro chronic antigenic stimulation. (c and d) Representative flow cytometry histograms (c) and bar plots (d) showing OPP incorporation in control, Larp4-KO, Nr4a1-KO, Nr4a2-KO and Nr4a1/2-KO Cas9+ T cells after 6 days of chronic antigenic stimulation (n = 4). P-values were calculated using two-sided unpaired Student’s t-tests. Data are shown as mean ± SEM. (e) Volcano plot showing mRNAs with differential TE between control and Larp4-KO TEX cells. LARP4-targeted mRNAs with significant differential TE (|log2FC| > 0.5, p-value < 0.01) are highlighted. (f) MSigDB pathway analysis of LARP4-targeted mRNAs with significantly downregulated TE after Larp4 knockout. The systematic name of each pathway in MSigDB is labeled. (g) Heatmap showing the TE of indicated genes in control and Larp4-KO in exhausted T cells after 8 days of in vitro chronic antigenic stimulation. (h) Bar plot showing the polysome / non-polysome ratio of indicated LARP4-targeted mRNAs and non-LARP4-targeted mRNA revealed by sucrose polysome profiling and qPCR (n = 2). The data represent two independent experiments (b, c, h) and three independent experiments (a).
