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. 2018 Oct 24;293(50):19365–19376. doi: 10.1074/jbc.RA118.004872

Figure 3.

Figure 3.

Trpt1 does not affect XBP1 or tRNATyr splicing even in the absence of RtcB in ES cells. A, RT-PCR analysis comparing spliced and unspliced XBP1 mRNA levels in RtcB cKO and Trpt1−/−, RtcB cKO cells. RtcB was depleted following a 3-day 4-OHT treatment. To induce ER stress, the cells were treated with tunicamycin (TUNI) for 4 h. The ladder is denoted with L. B, RT-qPCR analysis to quantify XBP1 splicing. Both spliced and total XBP1 levels were normalized to GAPDH. C, XBP1s protein levels were analyzed using chemiluminescence (top) and quantitative (bottom) Western blotting. The cells were treated with tunicamycin for 6 h or with DTT for 4 h followed by a 2-h recovery. Relative intensities of XBP1s are shown below the blot. D, RT-qPCR analysis of spliced and unspliced tRNATyr levels shows that spliced tRNATyr levels did not differ significantly between E8H2 and T10D6 cells. Error bars represent S.E. CTL, control.