Fig. 3

DHCR24 overexpression does not affect the levels of total cholesterol, but increases significantly the levels of NLG1 in the neuron-BV2 cell co-cultures. a Western blotting was used to detect the levels of total and phosphorylated forms of Akt, ERK1, and ERK2. DHCR24 overexpression had no significant effect on the levels of these proteins. However, the total Akt levels decreased, and phosphorylated Akt levels increased upon LPS and IFN-γ treatment. Phosphorylated ERK2 levels were significantly reduced after LPS and IFN-γ treatment. b Levels of total CREB are decreased in both control and DHCR24-overexpressing co-cultures upon LPS and IFN-γ treatment. Phosphorylation of CREB is increased upon LPS and IFN-γ treatment. Western blot samples in the figure are from the same membrane. c The overexpression of DHCR24 significantly increased NLG1 levels upon basal growth conditions in primary neuron-BV2 cell co-cultures. A similar trend towards increase was also observed in co-cultures overexpressing DHCR24 upon LPS and IFN-γ treatment. The assessment of phosphorylation status of cofilin-1 at Ser 3 did not show statically significant changes between DHCR24-overexpressing or control co-cultures after vehicle or LPS and IFN-γ treatments. The induction of neuroinflammation per se significantly reduced the phosphorylation cofilin-1 at Ser 3. d Total protein-normalized total cholesterol levels in control and DHCR24-overexpressing co-cultures upon vehicle and LPS and IFN-γ treatments do not show statistically significant changes between groups. Vehicle control was normalized to 100%. *p < 0.05, **p < 0.01, ***p < 0.001, mean ± SEM, n = 6 (a, b), n = 3 (c), and n = 7 (d)