Fig. 6.

VSIG4 promotes PDK2 expression through activating PI3K/Akt–STAT3. a Western blot of Akt and p-Akt^ser473 expression. b RAW264.7 cells were infected with different Vsig4 deletion constructs, cells were further treated with LPS (2 μg/ml), the expression of p-Akt^ser473 was analyzed by western blot. c Western blot of p-Akt^ser473 and PDK2 in VSIG4⁺RAW264.7 cells with mutation of Ser²⁷³, Ser²⁷⁶, Thr²⁷⁰, and Thr²⁷⁴ to Ala. VSIG4⁺RAW264.7 cells were treated with d the Akt inhibitor MK-2206, e the PI3K inhibitor Ly294002, and the expression of Akt, p-Akt^ser473, PDK2, and p-STAT3 was detected by western blot. f VSIG4⁺RAW264.7 cells were treated with the MK-2206, and the expression of STAT3/p-STAT3 was analyzed by western blot. g Western blot of p-STAT3/STAT3 and PDK2 in LPS-activated VSIG4⁺RAW264.7 cells followed with STAT3 inhibitor, S3I-201 (100 μM) treatment for 24 h. h Western blot of PDK2 in Stat3 silenced VSIG4⁺RAW264.7 cells. i The enrichment of p-STAT3 in Pdk2 gene promoter region was detected by ChIP-qPCR assay. j Human VSIG4⁺THP-1 cells were treated with microbeads-C3b (20 μg/ml) in the presence of LPS (2 μg/ml), and the expression of PDK2 was detected by western blot. Moreover, C3 ^−/− BMDMs were transfected to overexpress VSIG4, and cells were further treated with LPS (2 μg/ml) for an additional 3 h, and the expression of PDK2 was detected by western blot. Error bar, s.e.m. *p < 0.05, ***p < 0.0001 and NS, p > 0.05 (Student’s t-test). Data are representative of three independent experiments