Extended Data Fig. 3: Generation of LUHMES- and stem cell derived neurons.
From: TMEFF1 is a neuron-specific restriction factor for herpes simplex virus
a, Schematic illustration of the protocol for generation of LUHMES-derived neuron-like cells. b, Immunoblotting for TMEFF1 and vinculin in LUHMES neurons and effect of TMEFF1 sgRNA treatment. c, Confocal microscopy image of differentiated neurons stained with MAP2, betaIII-Tubulin, and DAPI, following the protocol shown in panel a (n = 3). d, LUHMES-derived neurons treated with TMEFF1 sgRNA #2 (Fig. 2b: TMEFF1 sgRNA #1) and infected with HSV-1-K64GFP (MOI 5) (n = 3 biological replicates pr time point). Cells were isolated after 8, 12, and 24 h, and examined by flow cytometry for GFP+ signal. Data are presented as % GFP+ cells. Two-tailed two-way ANOVA test followed by unpaired t-test comparing ctrl. with TMEFF1 sgRNA #2, comparing means, ±s.d. indicated by exact P-values (d).; P < 0.05 was considered statistically significant. e, Schematic illustration of the protocol for generation of ESC-derived cortical neurons. f, Immunoblotting for TMEFF1 in hESC-derived neurons and effect of TMEFF1 sgRNA treatment. g, Confocal microscopy image of differentiated neurons stained with MAP2, betaIII-Tubulin, and DAPI, following the protocol shown in panel e. Scale bar, 50 µm. h, Percentage of MAP2-positive cells, normalized to DAPI, in control and TMEFF1 sgRNA-transfected hESC-derived neurons. Immunoblots (b + f) and confocal imaging data (g) are representative of at least 3 independent experiments. Data is represented as mean ± s.d. (n = 5 biological replicates), and analysed using an unpaired two-tailed t-test. Exact P-values shown. P-values < 0.5 were considered significant. For gel source data, see Supplementary Fig. 1.
