Fig. 5: Analysis of MN and HB9-dependent IN genes upon nHOTAIRM1 KO.

A Expression profile, by qRT-PCR analysis, of ChAT and ISLET1 genes along the differentiation (from D0 to D12, x-axis) of WT or KO for nHOTAIRM1 spMNs (KO#2 clone). Data (means ± SEM) are expressed as arbitrary units, relative to ATP5O mRNA levels, as internal control. Expression levels at D0 were set as 1. N = 3 biological replicates; *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, two-tailed Student’s t-test, referred to D0 as the reference sample for statistical tests. B Quantification by immunostaining of CHAT + /ISLET1+ cells, expressed as percentage of the total number of cells, in WT and nHOTAIRM1 KO (KO#2) spMNs (D12). N = 3 groups of 60 cells randomly selected from 9 different fields, both for WT and KO, were analyzed. ***P ≤ 0.001. C qRT-PCR validation, in WT and nHOTAIRM1 KO (KO#2 clone) spMNs (D12) of the expression of HB9-dependent IN genes (indicated above each panel) significantly upregulated in HB9 KO MNs and nHOTAIRM1 KO spMNs compared to WT spMNs. Data (means ± SEM) are expressed in arbitrary units, relative to ATP5O mRNA levels, as internal control. For each target, the expression in WT spMNs was set as 1. N = 3 biological replicates; *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, two-tailed Student’s t test.