Figure 5.

TACI and Akt/mTOR/HIF-1α signaling were involved in BAFF priming of BV2 cells. BV2 cells were primed with 10ng/ml BAFF or vehicle for 24 h. (A) RNA was collected and the relative mRNA expression of TACI, BAFF-R, and BCMA, relative to GAPDH, are assessed in BV2 cells by quantitative real-time PCR. (B, C) The expression of BAFF receptors on the surface of microglia was detected by flow cytometry. BV2 cells are stained with primary antibodies for TACI, BAFF-R, and BCMA (solid lines without filling) or an isotype control Ab (solid lines filled with gray). (B) The graph represents the relative fluorescence intensity of the primary antibody subtracting its isotype control antibody. (D) 6 days after BV2 cells were primed by BAFF or vehicle, whole-cell extracts were collected and levels of Akt, p-Akt, mTOR, p-mTOR, and HIF-1α were detected by western blot. Densitometric quantification of p-Akt/Akt (E), p-mTOR/mTOR (F), and HIF-1α/tubulin (G) in (D). Data are mean ± SEM (n = 9); Kruskal-Wallis test with Turkey post hoc test; *p < 0.05, **p < 0.01 compared with CTL.