Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: CCL11 (Eotaxin) Induces CCR3-Dependent Smooth Muscle Cell Migration

doi: 10.1161/01.atv.0000131654.90788.f5

Figure Lengend Snippet: Figure 3. CCL11 induces mouse SMC chemotaxis. A, Mouse CCL11 in 0.3% FCS at the concentrations indicated (ng/mL) were placed in the bottom wells of a modified Boyden Cham- ber. 104 SMCs were placed in the top wells. The chamber was incubated for 6 hours at 37°C and then analyzed for migration as described in Materials and Methods. For comparison, recom- binant PDGF-BB (1 ng/mL) or 10% FBS were also used as che- moattractants. 0.3% serum was used as negative control. *P0.005 compared with 0 CCL11. B, Checkerboard analysis in which CCL11 (C; 100 ng/mL) was placed in the bottom, top, or both wells as indicated. *P0.005 compared with 0.3% FCS (–). C, SMC were incubated with antibodies against CCR3 (anti- CCR3), CCR2 (anti-CCR2) (1 g/mL), or irrelevant IgG for 30 minutes at 37°C before being placed in the Boyden chamber. Results are expressed as SMC/hpf SEM and represent tripli- cate experiments *P0.005 compared with 0.3% FCS. #P0.05 compared with 0.3% FCS.

Article Snippet: Recombinant mouse CCL11 (420-ME; R & D Systems, Minneapolis, MN) was added to the bottom chamber in 0.3% FBS and DMEM.

Techniques: Chemotaxis Assay, Incubation, Migration, Comparison, Negative Control

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: CCL11 (Eotaxin) Induces CCR3-Dependent Smooth Muscle Cell Migration

doi: 10.1161/01.atv.0000131654.90788.f5

Figure Lengend Snippet: Figure 5. CCL11 does not modulate SMC proliferation. SMCs were plated in 35-mm wells and treated with CCL11 (10 ng/mL or 100 ng.ml) PDGF (1 ng/mL), CCL11 (100 ng/mL) PDGF (1 ng/mL), 10% FCS, or 0.3% FCS. Duplicate plates were counted on the days indicated. The medium, with fresh growth factors, was replaced every 48 hours. Results are expressed as cell number SEM and represent the average of duplicate experiments. Figure 4. CCL11 induces mouse SMC migration. SMCs were grown to confluence and then incubated in DMEM 0.1% BSA for 24 hours. Linear wounds were made in each plate and SMCs were then incubated with DMEM 0.1% BSA alone (con- trol) or in the presence of CCL11 (concentration in ng/mL indi- cated on the X axis). In some experiments, SMCs were preincu- bated for 1 hour with anti-CCR3 (CCR3 Ab) or an irrelevant antibody (IgG) before wounding and treatment with 10 ng/mL CCRL11. The wound area was measured immediately after wounding and at 24 hours. Migration is expressed as the % decrease in wound area in 24 hours compared with control. Experiments were done in duplicate, with 3 to 5 plates used for each condition. *P0.001, 10 ng/mL CCL11 compared with control; **P0.001, 10 ng/mL eotaxin CCR3 Ab compared with 10 ng/mL eotaxin alone; #NS, 10 ng/mL eotaxin IgG compared with 10 ng/mL eotaxin alone.

Article Snippet: Recombinant mouse CCL11 (420-ME; R & D Systems, Minneapolis, MN) was added to the bottom chamber in 0.3% FBS and DMEM.

Techniques: Migration, Incubation, Concentration Assay, Control

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: CCL11 (Eotaxin) Induces CCR3-Dependent Smooth Muscle Cell Migration

doi: 10.1161/01.atv.0000131654.90788.f5

Figure Lengend Snippet: Figure 6. Expression of CCR3 antigen in injured mouse femoral arteries. Samples were obtained from a previous study in which C57Bl/6 mice had undergone wire-induced femoral arterial injury.39 Sections from uninjured arteries (A, B), arteries harvested 5 days (C, D), 7 days (E, F), and 28 days after injury (G through J) were stained with antibodies to CCR 3 (left) or CCL11 (right). Magnifica- tion40 for A, B, E, F, G, and H and 200 for C, D, I, and J. Sections are representative of studies done on 3 different animals.

Article Snippet: Recombinant mouse CCL11 (420-ME; R & D Systems, Minneapolis, MN) was added to the bottom chamber in 0.3% FBS and DMEM.

Techniques: Expressing, Staining

Journal: mBio

Article Title: Host cysteine proteases promote the severity of catheter-associated urinary tract infection and kidney fibrosis

doi: 10.1128/mbio.02161-25

Figure Lengend Snippet: Eosinophils play a protective role in CAUTI. At 2 dpi, single cells were isolated from the bladder mucosa of CAUTI mice. The percentages of eosinophils and neutrophils were measured by flow cytometry to assess the effects of E64 treatment ( A ) or eotaxin ( B ) as compared to mock. Total CFU recovered 2 dpi were quantified for catheter, bladder, and kidney for treatment by eotaxin ( C–E ), by a-IL-5 ( F–H ), or by CCR3 antagonist SB328437 ( I–K ). Each symbol represents an individual mouse, and symbols touching the dashed lines indicate values below the limit of detection (LOD, 40 CFU). Horizontal lines indicate mean values; error bars represent SEM. All data are from ≥3 biological replicates. Statistical comparisons were performed using a two-tailed unpaired Student’s t-test. * P < 0.05. Normality was assessed using the Shapiro-Wilk test.

Article Snippet: For drug therapy, E64 (Roche, 10874523001), anti-mouse IL-5 (BD Pharmingen, 554393), SB 328437 (CCR3 antagonist, Sigma, 247580-43-4), and mouse eotaxin (R&D systems, 420-ME) were dissolved in PBS according to the manufacturers’ recommendations to achieve the concentration stated in the text.

Techniques: Isolation, Flow Cytometry, Two Tailed Test