Kir7.1 regulates chemotaxis and GPCR signaling of human neutrophils. (A) Under-agarose migration assay of dHL-60 cells treated with 50 μM VU590 or DMSO on BSA- or collagen-coated glass-bottom dishes toward indicated chemoattractants. (B and C) Quantification of forward migration index (FMI) toward the chemokine gradient and velocity of cells. n > 10 in each group. Data present mean ± SD, representative of three independent experiments. (D) Under-agarose random migration velocity of dHL-60 cells treated with 50 μM VU590 or DMSO on BSA-coated glass-bottom dishes in the presence or absence of uniform fMLP. n = 20 in each group. Data present mean ± SD, pooled from three independent experiments. (E) Whole-cell patch-clamp measurement of dHL-60 cells treated with VU590 or KCl with or without fMLP activation. n > 10 in each group. Data present mean ± SD, representative of three independent experiments. (F) Illustration of how Kir regulates MP and GPCR signaling. (G) Quantification of intracellular calcium after fMLP treatment of dHL-60 cells treated with VU590 or DMSO. Data present mean ± SD, representative of three independent experiments. (H and I) Immunoblot and normalized quantification of time-course PAK activation after fMLP treatment of dHL-60 cells treated with DMSO, VU590, or KCl. Data present mean ± SD from three independent experiments. (B, C, and G) Mann–Whitney test. (D, E, and I) Multiple comparisons and one-way ANOVA. Source data are available for this figure: SourceData F5.
Figure 5.

Kir7.1 regulates chemotaxis and GPCR signaling of human neutrophils. (A) Under-agarose migration assay of dHL-60 cells treated with 50 μM VU590 or DMSO on BSA- or collagen-coated glass-bottom dishes toward indicated chemoattractants. (B and C) Quantification of forward migration index (FMI) toward the chemokine gradient and velocity of cells. n > 10 in each group. Data present mean ± SD, representative of three independent experiments. (D) Under-agarose random migration velocity of dHL-60 cells treated with 50 μM VU590 or DMSO on BSA-coated glass-bottom dishes in the presence or absence of uniform fMLP. n = 20 in each group. Data present mean ± SD, pooled from three independent experiments. (E) Whole-cell patch-clamp measurement of dHL-60 cells treated with VU590 or KCl with or without fMLP activation. n > 10 in each group. Data present mean ± SD, representative of three independent experiments. (F) Illustration of how Kir regulates MP and GPCR signaling. (G) Quantification of intracellular calcium after fMLP treatment of dHL-60 cells treated with VU590 or DMSO. Data present mean ± SD, representative of three independent experiments. (H and I) Immunoblot and normalized quantification of time-course PAK activation after fMLP treatment of dHL-60 cells treated with DMSO, VU590, or KCl. Data present mean ± SD from three independent experiments. (B, C, and G) Mann–Whitney test. (D, E, and I) Multiple comparisons and one-way ANOVA. Source data are available for this figure: SourceData F5.

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