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. 2020 Jan 13;39(4):e100574. doi: 10.15252/embj.2018100574

Figure 3. Decreased expression of C9ORF72 synergizes DPR protein toxicity.

Figure 3

  1. Cell viability (TO‐PRO‐3 FACS staining) of GT1‐7 neuronal cells transfected for 24 h with either wild‐type or indicated mutant constructs containing either 80 G4C2 repeats or 100 C4G2 repeats embedded in sense or antisense C9ORF72 fused to the GFP in the GA, PG, or GR frame.
  2. Immunoblotting against the GFP, endogenous C9ORF72, or the GAPDH of proteins extracted from Neuro2A cells transfected for 24 h with 80 G4C2 repeats embedded in the human sense C9ORF72 sequence fused to the GFP in the GA frame and with either a control siRNA or a siRNA targeting C9orf72 mRNA.
  3. Immunoblotting against the HA‐tag, endogenous C9ORF72, or GAPDH of proteins extracted from Neuro2A cells transfected for 24 h with 100 C4G2 repeats embedded in the human antisense C9ORF72 sequence fused to a HA‐tag in the PG frame and with either a control siRNA or a siRNA targeting C9orf72 mRNA or treated with bafilomycin A1 for 15 h.
  4. Immunoblotting against the HA‐tag, endogenous C9ORF72, or GAPDH of proteins extracted from Neuro2A cells transfected for 24 h with 80 G4C2 repeats embedded in the human sense C9ORF72 sequence fused to a HA‐tag in the GR frame and with either a control siRNA or a siRNA targeting C9orf72 mRNA or treated with bafilomycin A1 for 15 h.
  5. Cell viability (TO‐PRO‐3 FACS staining) of GT1‐7 neuronal cells co‐transfected for 24 h with either a control siRNA or a siRNA targeting C9orf72 mRNA and wild‐type or mutant constructs containing either 80 G4C2 repeats or 100 C4G2 repeats embedded in sense or antisense C9ORF72 fused to the GFP in the GA, PG, or GR frame.
Data information: Error bars indicate s.e.m. Student's t‐test, *P < 0.05, **P < 0.01, and ***P < 0.001. n = 5 independent transfection.