Fig. 3

ER stress genes up-regulated in SHSY-5Y cells under MANF treatment. a Differential gene expression between 6-OHDA treatment group and 6-OHDA + MANF treatment group based on RNA-seq analyses, 2027 genes up-regulated; 2736 genes down-regulated; b: Gene ontology analysis of the top 26 biological processes associated with up regulated gene expression in the 6-OHDA + MANF treatment group relative to 6-OHDA treatment group (ranked by q-value). c, d: RNA-seq with whole gene profiling was performed to detect the differentially expressed genes in 6-OHDA + MANF cultures vs. 6-OHDA cultures in SHSY-5Y cells. These genes including HSP70, GRP78, MAPK, ATF6, ATF4 and xbp-1. e: Real-time PCR quantitation of HSP70, GRP78, xbp-1, ATF-4, ATF-6, MAPK mRNA levels in control, 6-OHDA and 6-OHDA + MANF treatment group (mean ± SEM, n = 5). The relative mRNA expression levels of HSP70, GRP78, xbp-1, ATF-4, ATF-6, MAPK in SHSY-5Y cells were normalized to those of GAPDH. (*p < 0.05; one-way ANOVA with Bonferroni post-tests). f: A representative Westernblot result shows the expression of phosphorylated PERK and PERK in control SHSY-5Y cells, SHSY-5Y cells treated with 250 μM of 6-OHDA; SHSY-5Y cells treated with 4 μg/mL of MANF; and SHSY-5Y cells treated with 250 μM 6-OHDA +4 μg/mL MANF at 5 min