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1.
FIG 5

FIG 5. From: Mitogen-Activated Protein Kinase 2 Signaling Shapes Macrophage Plasticity in Aggregatibacter actinomycetemcomitans-Induced Bone Loss.

MK2 differentially regulates chemokines CCL3 and CCL4. (A) ELISA quantification of CCL3 in air pouch exudate from A. actinomycetemcomitans-treated mice (n = 6 to 10). (B) ELISA quantification of CCL4 in air pouch exudate from A. actinomycetemcomitans-treated mice (n = 6 to 10). Data are expressed as means ± SEMs compared to the results for the WT control groups. *, P ≤ 0.05.

Bethany A. Herbert, et al. Infect Immun. 2017 Jan;85(1):e00552-16.
2.
FIG 4

FIG 4. From: Mitogen-Activated Protein Kinase 2 Signaling Shapes Macrophage Plasticity in Aggregatibacter actinomycetemcomitans-Induced Bone Loss.

MK2 signaling does not regulate circulating monocytes during A. actinomycetemcomitans pathogenesis. (A) Representative flow cytometry plots of peripheral blood CD11b+ CCR1+, CD11b+ CCR2+, and CD11b+ CCR5+ cells from Mk2+/+ and Mk2−/− mice treated with PBS vehicle or A. actinomycetemcomitans (A.a.) for 3 days. Q1 to Q4, quadrants 1 to 4, respectively. (B) Quantification of percent positive CD11b+ CCR1+, CD11b+ CCR2+, and CD11b+ CCR5+ cells from Mk2+/+ and Mk2−/− mice treated with PBS vehicle or A. actinomycetemcomitans for 3 days (n = 4). Data are expressed as means ± SEMs. *, P ≤ 0.05.

Bethany A. Herbert, et al. Infect Immun. 2017 Jan;85(1):e00552-16.
3.
FIG 2

FIG 2. From: Mitogen-Activated Protein Kinase 2 Signaling Shapes Macrophage Plasticity in Aggregatibacter actinomycetemcomitans-Induced Bone Loss.

Aggregatibacter actinomycetemcomitans stimulates MK2 signaling in macrophages and in vivo. (A) A representative immunoblot of A. actinomycetemcomitans (A.a.) treatment (tx) stimulating p38 and MK2 phosphorylation in murine BMDMs (n = 2). (B) A representative immunoblot of calvarial tissue protein treated with A. actinomycetemcomitans for 3 days. (C) Densitometric analysis of calvarial tissue proteins in which the protein levels were normalized to the levels of GAPDH (n = 3 to 5). Data are expressed as means ± SEMs. *, P ≤ 0.05.

Bethany A. Herbert, et al. Infect Immun. 2017 Jan;85(1):e00552-16.
4.
FIG 3

FIG 3. From: Mitogen-Activated Protein Kinase 2 Signaling Shapes Macrophage Plasticity in Aggregatibacter actinomycetemcomitans-Induced Bone Loss.

MK2 regulates chemokines in A. actinomycetemcomitans-challenged macrophages and in vivo. (A) RT-qPCR of Ccl3 and Ccl4 gene expression of Mk2+/+ and Mk2−/− BMDMs stimulated with 10 CFU of A. actinomycetemcomitans (A.a.) per cell or untreated control Mk2+/+ and Mk2−/− BMDMs (n = 3 to 4). (B) RT-qPCR of Ccl3 (left) and Ccl4 (right) gene expression in mouse calvarial tissue challenged with A. actinomycetemcomitans or PBS vehicle for 3 days (n = 3 to 7). (C) Multiplex assay of mouse tissue chemokines after 3 days of A. actinomycetemcomitans treatment (n = 4). (D) Multiplex assay of mouse peripheral blood chemokines after 3 days of A. actinomycetemcomitans treatment (n = 4). Data are expressed as means ± SEMs. *, P ≤ 0.05; **, P ≤ 0.01.

Bethany A. Herbert, et al. Infect Immun. 2017 Jan;85(1):e00552-16.
5.
FIG 1

FIG 1. From: Mitogen-Activated Protein Kinase 2 Signaling Shapes Macrophage Plasticity in Aggregatibacter actinomycetemcomitans-Induced Bone Loss.

MK2 signaling regulates A. actinomycetemcomitans-induced inflammatory infiltrate. (A) Representative ×40-magnification images of Mk2+/+ and Mk2−/− mouse calvaria, including the epithelium (E), calvarium (C), and brain (B), treated with PBS vehicle or A. actinomycetemcomitans (A.a.) for 3 or 5 days. Blue scale bars, 100 μm. (B) Inflammatory infiltrate scores for Mk2+/+ and Mk2−/− mice treated with PBS vehicle or A. actinomycetemcomitans for 3 days (top) and 5 days (bottom). (C) Representative ×200-magnification images obtained after F4/80 staining of A. actinomycetemcomitans-treated Mk2+/+ and Mk2−/− mice for 3 days and Visiopharm analysis. (D) F4/80-positive (F4/80+) area of Mk2+/+ and Mk2−/− mice treated with PBS vehicle or A. actinomycetemcomitans for 3 and 5 days. (E) Analysis of macrophage marker RNA in mouse calvarial tissue treated for 3 days by use of the NanoString Technologies immunology panel (n = 3 to 5). Data are expressed as means ± SEMs. *, P ≤ 0.05; **, P ≤ 0.01.

Bethany A. Herbert, et al. Infect Immun. 2017 Jan;85(1):e00552-16.
6.
FIG 6

FIG 6. From: Mitogen-Activated Protein Kinase 2 Signaling Shapes Macrophage Plasticity in Aggregatibacter actinomycetemcomitans-Induced Bone Loss.

A. actinomycetemcomitans induces murine calvarial bone loss through MK2. (A) Representative three-dimensional reconstruction of μCT results for mouse calvaria. Red arrows, areas of calvarial resorption that have been magnified 6×. (B) Calvaria resorption pit enumeration for A. actinomycetemcomitans-treated mice (n = 3 to 5). (C) Representative ×200-magnification images of mouse calvaria for TRAP staining for osteoclasts. Red arrows, osteoclasts in contact with bone surface. (D) Results of analysis of tnfsf11 (RANKL) RNA counts from mouse calvarial tissue treated for 3 days (n = 3 to 5) by use of the NanoString Technologies immunology panel. (E) Enumeration of the number of osteoclasts (OC) per calvarial bone perimeter (B.Pm.) for mice treated for 3 days and 5 days (n = 3 to 5). Data are expressed as means ± SEMs. *, P ≤ 0.05; **, P ≤ 0.01.

Bethany A. Herbert, et al. Infect Immun. 2017 Jan;85(1):e00552-16.

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