Fig 1: Transcriptional Dynamics and BMP Signaling across Mouse In Vitro Neurogenesis(A) Experimental design. Passage 1 (PS1) NSCs were passaged into N2 + different FGF2 doses (PS2). Differentiation was induced by FGF2 withdrawal at DIV2. Neurotrophins (NTs) were added from DIV4.(B) Six of 11 GWCoGAPS patterns shown in Figure S1. p values indicate the enrichment of GO categories in each pattern.(C–E) Top: expression dynamics of selected gene clusters annotated by “BMP receptor signaling” from NCI (full dendrogram in Figure S1), correlating with BMP7 (C), BMP4 (D), and BMP6 (E). BMP7-related genes were rapidly repressed in low FGF2. BMP4-related gene levels increased at the initiation of differentiation. BMP6-related gene levels increased at late time points. Bottom: gene weights in each GWCoGAPS pattern for the same upper genes.(F) Immunofluorescence images of SYN1 puncta in neurons cultured over astrocytes (i) at different FGF2 doses, or (ii) at 1 ng/mL FGF2 + LDN (100 nM). Scale bar: 10 µm. (iii) SYN1 and HOM1 puncta counts per 100 mm of neurite length for each condition, assessed by high-throughput image analysis. Means ± SDs, t test, n > 5 fields per measurement. LDN or BMP4 were added 12 h after NSC plating and withdrawn with FGF2 at DIV2 (see A).