Fig. 6: PolyGR induces TGFB1 followed by its target gene COL6A1 in i3Neurons.
a, The top ten IPA-predicted upstream regulators in (GR)400 spinal cord (left panel), (PR)400 spinal cord (center panel) and C9orf72 iPS cell-derived motor neurons (right panel). WT (n = 4 mice), (GR)400 (n = 5 mice), (PR)400 (n = 6 mice); two-sided Welch’s t-test with 5% FDR multiple-correction. b, Boxplot of residual gene expression for TGFB1. The boxplot midline represents the median, the box represents the interquartile range (25th and 75th percentiles) and the whiskers extend to the highest (upper whisker) or lowest (lower whisker) value, but no more than the interquartile range multiplied by 1.5. To determine differentially expressed genes, a linear regression model (two-sided) was used and P values were adjusted for multiple comparisons using the Benjamini–Hochberg FDR procedure. C9orf72 FTLD/MND n = 34, non-C9orf72 FTLD/MND n = 44, controls n = 24. c, Schematic workflow of i3Neuron doxycycline-induced differentiation at DIV 0 and transduction with lentiviruses expressing (GR)50, or GFP at DIV 3. d, Live-cell Incucyte confluency quantification in (GR)50, or GFP-treated i3Neurons. Graph, mean ± s.e.m., n = 3 independent biological replicates, two-way ANOVA, Bonferroni’s multiple comparison. e, qPCR analysis of TGFB1 transcript levels normalized to GAPDH at DIV 5 (left panel) and DIV 7 (right panel) in (GR)50, or GFP-treated i3Neurons. Graph, mean ± s.e.m., n = 3 independent biological replicates, one-way ANOVA, Bonferroni’s multiple comparison. f, qPCR analysis of COL6A1 transcript levels normalized to GAPDH at DIV 5 (left panel) and DIV 7 (right panel) in (GR)50, or GFP-treated i3Neurons. Graph, mean ± s.e.m., n = 3 independent biological replicates, one-way ANOVA, Bonferroni’s multiple comparison. NS denotes P > 0.05. FTLD, frontotemporal lobar degeneration.
