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. 2018 Apr 9;13(4):e0194310. doi: 10.1371/journal.pone.0194310

Fig 3. Effect of ASB7 knockdown on UPR markers and pathways under ER stress.

Fig 3

(A) HeLa cells were transiently transfected with 2 independent ASB7 siRNA and a scrambled control (SC). After 2 days, levels of endogenous ASB7 mRNA and protein levels were measured by qRT‐PCR and immunoblotting and normalized relative to the housekeeping gene, 18S rRNA and β-actin (mean ± SEM, n = 2). (B) HeLa cells were transiently transfected pooling 2 independent ASB7 siRNAs (gray bars) and compared with scrambled siRNA as control (SC) (black bars). After 2 days, cells were cultured with DTT (2 mM) and Tun (20 μg/ml) for 3 h then total RNA was extracted. Relative levels of mRNAs encoding CHOP, GRP78, PERK, IRE-1α, ATF6 and spliced XBP-1 ratio were compared by qRT-PCR and displayed as ratios relative to a housekeeping gene (18S rRNA). “*” denotes significance with respect to scramble control. Error bars represent mean ± SEM. *** p≤0.001; **: p≤0.01; *: p≤0.05 calculated using one-way ANOVA.