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. Author manuscript; available in PMC: 2020 Aug 18.
Published in final edited form as: Cell Rep. 2020 Jul 14;32(2):107895. doi: 10.1016/j.celrep.2020.107895

Figure 2. The V-ATPase Is Required for NMD.

Figure 2.

(A) Flow cytometry analysis of EGFPPTC-231 cells after lentiviral delivery of Cas9 and NT or ATP6V0C-targeting sgRNAs.

(B and C) qRT-PCR analysis of EGFPPTC-231 (B) or EGFPPTC-35 mRNA (C) in reporter cell lines after lentiviral delivery of Cas9 and the indicated sgRNAs. All mRNA expression levels shown in this figure were first normalized to GAPDH expression and then normalized to expression level in sgNT1-infected cells.

(D) Analysis of EGFPPTC-231 mRNA stability in tet-OFF-EGFPPTC-231 cells after lentiviral delivery of Cas9 and the indicated sgRNAs. Cells were treated with 1 μg/mL doxycycline, and the EGFP mRNA level relative to GAPDH abundance at the indicated time points was determined by qRT-PCR.

(E and F) qRT-PCR analysis of endogenous NMD targets (E) or PTC+ and PTC− isoforms of SRSF2 (F) in HCT116 cells after lentiviral delivery of Cas9 and the indicated sgRNAs.

Data are represented as mean ± SD (n = 3 biological replicates for all qRT-PCR experiments). *p < 0.05, **p < 0.01, ***p < 0.001; calculated by Student’s t test. See also Figure S2.