Fig. 6: RPS29 maintains STMN2 translation and protein expression. | Communications Biology

Fig. 6: RPS29 maintains STMN2 translation and protein expression.

From: Machine learning-based proteomics profiling of ALS identifies downregulation of RPS29 that maintains protein homeostasis and STMN2 level

Fig. 6

Bar plots showing the gene set enrichment analysis (GSEA) of RNA-seq a and Ribo-seq b after RPS29 knockdown in SY5Y. NES, normalized enrichment score. c Heatmap showing the translation levels of key ALS-related proteins. Immunoblots d and quantification results e showing decreased expression levels of ALS-related proteins in SH-SY5Y after RPS29 knockdown. Each dot represents one biological replicate (n = 3). Data are presented as mean ± SD. The p value was calculated using a two-tailed t-test. Immunoblots f and quantification results g showing increased levels of poly-GA produced by repeat-associated non-AUG (RAN) translation of C9orf72 repeats after RPS29 knockdown in HEK-293T cells. Each dot represents one biological replicate (n = 3). Data are presented as mean ± SD. The p value was calculated using a two-tailed t-test. h Images (left) showing increased levels of poly-GA produced by repeat-associated non-AUG (RAN) translation of C9orf72 repeats after RPS29 knockdown in HEK-293T cells. The cell proportion (right) was obtained by dividing the DPR cells in each field of view by all cells in the field of view (n = 10). Data are presented as mean ± SD. The p value was calculated using a two-tailed t-test. i Relative cell viability of motor neuron with shRNA-mediated knockdown of RPS29. Each dot represents a biological replicate (n = 3). Data are presented as mean ± SD. The p value was calculated using a two-tailed t-test. j Images (left) and quantification results (right) showing decreased levels of STMN2 in the axon of motor neurons after RPS29 knockdown. Relative STMN2 expression level was obtained by dividing the mean intensity of the axons by the mean intensity of the soma in each neuron (n = 28). Data are presented as mean ± SD. The p value was calculated using a two-tailed t-test. TUJ1, a neuron marker. k Relative cell viability of SH-SY5Y and motor neuron with shRNA-mediated knockdown of RPS29 and RPS29 overexpression. Each dot represents one biological replicate (n = 3). All data are representative of at least three independent experiments. Data are presented as mean ± SD. The p value was calculated using one-way ANOVA. *p < 0.05; **p < 0.01; ****p < 0.0001; ns not significant.

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