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. 2021 Nov 20;23:582–592. doi: 10.1016/j.omto.2021.11.012

Figure 2.

Figure 2

Restoring protein expression of LDLR and OAS1 decreased titers

(A) LDLR protein expression in parental 293T, LDLR−/− 293T, LDLR−/− 293T transduced with Lenti/GFP, and LDLR−/− 293T transduced with Lenti/LDLR measured by flow cytometry. LDLR−/− cells were transduced with an LV encoding LDLR to restore its expression, or an LV encoding GFP as a transduction control. (B) Percentage of LDLR+ cells in parental, LDLR−/−, and LDLR restored cells measured by flow cytometry. (C) Titers of Lenti/βAS3-FB packaged in parental 293T, LDLR−/− 293T, LDLR−/− 293T transduced with Lenti/GFP, and LDLR−/− 293T transduced with Lenti/LDLR. Because Lenti/LDLR carries an ires-GFP cassette, all transduced cells were sorted for the GFP+ population. The sorted GFP+ cells were expanded for VCN analysis by ddPCR and packaging (n = 9 dishes of identical cultures from three independent experiments; bars represent mean with SD; unpaired t test, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001). (D) OAS1 protein expression in parental 293T, OAS1−/− 293T, OAS1−/− 293T transduced with Lenti/GFP, and OAS1−/− 293T transduced with Lenti/OAS1 measured by western blot. (E) Titers of Lenti/βAS3-FB packaged in parental 293T, OAS1−/− 293T, OAS1−/− 293T transduced with Lenti/GFP, and OAS1−/− 293T transduced with Lenti/OAS1 (n = 12 dishes of identical cultures from four independent experiments; bars represent mean with SD; unpaired t test, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001).