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. 2019 May 24;26:45–56. doi: 10.1016/j.molmet.2019.05.009

Figure 5.

Figure 5

The RNA helicase DDX3X is required for efficient translation of JUND mRNA. (A) RT-qPCR for DHX9, DDX1, and DDX3X and (B) western blot for DDX3X show a significant depletion of these RNA helicases using CRISPR-Cas9 in Min6 cells (n = 3). (C) Representative western blot depicting JUND levels in Min6 cells transduced with lentivirus to deplete the indicated RNA helicase using CRISPR-Cas9 and cultured in glucolipotoxic conditions for 30hrs (n = 3). (D) Assessment of JUND mRNA abundance in Min6 cells by RT-qPCR after CRISPR-mediated depletion of DDX3X and culturing in glucolipotoxic conditions for 30hrs (n = 3). (E) Decreased ribosome occupancy of JUND in GFP-RPL10a Min6 cells after depletion of DDX3X by CRISPR-Cas9 and 30 h of glucolipotoxic conditions, as determined by TRAP followed by RT-qPCR (n = 3). * = p < 0.05 and ns = not significant. P values were calculated by two-way ANOVA except in (B) in which an unpaired two-tailed Student's t-tests was used. In (D), * denotes significance compared to ROSA26 group of the same treatment unless otherwise noted. For western blot images of JUND, arrows denote two bands for JUND and * denotes a non-specific band. Otherwise, * = p < 0.05. Data are presented as mean ± standard error of the mean unless otherwise noted.