Fig. 5.

ALKBH5 selectively enhances migration-promoting molecules but suppresses migration-inhibitory genes in neutrophils. a, b qRT-PCR of the mRNA levels of indicated genes in peritoneal neutrophils isolated from Alkbh5-deficient mice (Alkbh5^–/–) and WT littermates (Alkbh5^+/+) at 12 h (a) or 36 h (b) after mild CLP (n = 5). c qRT-PCR of the mRNA levels of indicated genes in ALKBH5-deficient (ALKBH5^–/–) and WT (ALKBH5^+/+) dHL-60 cells infected with E.coli as indicated times (n = 5). d qRT-PCR of the mRNA levels (relative to 0 h) of indicated genes in WT dHL-60 cells infected with E.coli as indicated times (n = 5). e FACS analysis of the cell-surface protein-level expression of CXCR2 on ALKBH5^+/+ and ALKBH5^–/– dHL-60 cells infected with E.coli for 6 h. f FACS analysis of the cell-surface protein-level expression of CXCR2 on WT dHL-60 cells infected with E.coli or uninfected as indicated. g Quantification of CXCR2 MFI on indicated dHL-60 cells as in (e) and (f) (n = 3). MFI mean fluorescence intensity. h, i Immunoblot analysis of the protein expression levels of indicated genes in blood neutrophils from Alkbh5-deficient mice (Alkbh5^–/–) and WT littermates (Alkbh5^+/+) at 12 h (h) or 36 h (i) after mild CLP. All data are mean ± SEM of biologically independent samples. qRT-PCR data were normalized to Gapdh (a, b) or GAPDH (c, d) expression. Data are representative of 3 independent experiments with similar results [(e, f) and (h, i)]. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001; ns not significant. Two-tailed unpaired Student’s t test [(a–d) and (g)]