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. 2021 Feb 3;11:582070. doi: 10.3389/fmicb.2020.582070

FIGURE 2.

FIGURE 2

LyzMcre KLF4 knockout develops in mature but not premature murine PMNs and reduces the killing of S. pneumoniae in murine blood-derived PMNs. PMNs were isolated from bone marrow (A) or blood (B) of control (KLF4+/+, black bars), heterozygous myeloid KLF4 knockout (KLF+/−, white bars), and homozygous myeloid KLF4 knockout (KLF4− /−, gray bars) mice and stimulated with R6× pneumococci (MOI 1 for 6 h). Cell lysates were collected after stimulation and analyzed for KLF4 expression using Western blotting. β-Actin confirmed equal protein loading. The densitometries of the KLF4 and β-actin bands were quantified using an Odyssey 2.0 infrared imaging system. The ratios of the KLF4 and β-actin densitometries were calculated and are shown as the fold change of induction to control (KLF4+/+) PMNs. Quantifications show the mean with standard deviation of three independent experiments. Blood-derived PMNs from control (KLF4+/+, black lines) and KLF4 knockout (KLF4− /−, gray lines) mice were stimulated with opsonized D39 (C) or opsonized R6× pneumococci (D) (MOI 100 for 1 and 3 h, input, dashed lines) for the CFU assay. Graphs show mean with standard deviation of CFU in percent of input (set to 100%) of three independent experiments. Statistics: Kruskal–Wallis test with Dunn multiple-comparisons test (A,B) or two-way ANOVA with Bonferroni post hoc test (C,D). ****p < 0.0001; *p < 0.05; n.s., not significant.