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. 2023 Mar 9;13:3934. doi: 10.1038/s41598-023-31141-6

Figure 3.

Figure 3

Generation and validation of CHOP reporter clonal cell lines. (A) RFP was introduced downstream the CHOP gene, resulting in a shift in the PCR amplicon. Red arrows indicate position of primers. (B) Time-course showing increase in RFP expression in iNeurons derived from the four clonal cell lines, following treatment with 100 nM Tun. The two homozygote lines (orange bars) show approximately two-fold higher median fluorescent intensity (MFI) when compared to the heterozygote lines (purple bars). (C) The Tun response (100 nM Tun-treated minus Unt MFI) is reduced with increasing concentrations of GSK2606414, a known Tun inhibitor. (D) ISRIB, another inhibitor of the pathway, reduced the Tun response. Data are shown as ± SEM, n > 3. One-way ANOVA with Dunnett’s multiple comparisons test. **p < 0.01, ***p < 0.001 and ****p < 0.0001 compared to 100 nM Tun only control (blue bar).