Fig. S2.

Effect of LDL treatment on proliferation and differentiation potential of precursor cells in vitro. (A) Cell proliferation was estimated by quantifying the number of primary neurospheres after 12 days exposure to LDL (0–200 μg/mL). (B-D) Following, neurospheres were collected and placed on PDL-laminin coated slides and allowed to differentiate for 7 days in absence of growth factors and no further LDL addition. Data in (A) were log2 transformed to visualize concentration-dependent response to LDL. Decrease of 1 indicates drop in neurosphere numbers by half. (D) Immunostaining against βIII- tubulin (red) and GFAP (green) identified neurons and astrocytes, respectively. LDL treatment decreased neuronal differentiation (B) and concomitantly increased proportion of astrocytes (C). *p < 0.05. Neurosphere numbers were analyzed with one-way ANOVA followed by Dunnett’s post hoc. The ratios of bIII-tubulin- and GFAP-positive cells were analyzed using generalized linear model with logit link function. Likelihood ratio test against the model omitting treatment showed significant effect of LDL on differentiation potential of NPCs [LRT, Chisq(4) = 195.23; p < 2.2e-16]. Post-hoc comparisons were performed using Dunnett’s test. The significance of the difference between control and 100 mg treatment group could not be estimated due to lack of variation in the 100 mg group (there were no neurons). N = 4 preparations (A); N = 84 neurospheres in two experimental batches (B). Scale bar = 20 μm. Data are presented as means from experiments ± SEM.