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Results: 7

1.
Figure 7

Figure 7. From: Caveolin-1 plays a critical role in host immunity against Klebsiella pneumoniae by regulating STAT5 and Akt activity.

Schematic diagram describing the proposed GSK3β–β-catenin–Akt axis involved in the dysregulated proinflammatory response. Cav1 knockout decreases β–catenin while increases GSK-3β expression, resulting in a decrease in Akt levels. This also downregulates STAT5 with and leads to decreased nuclear translocation, which may impact on production of cytokines such as TNF-α, IL-12a and IL-6, and infection outcome.

Qiang Guo, et al. Eur J Immunol. 2012 June;42(6):1500-1511.
2.
Figure 3

Figure 3. From: Caveolin-1 plays a critical role in host immunity against Klebsiella pneumoniae by regulating STAT5 and Akt activity.

Cav1 deficiency altered the inflammatory responses to K. pneumoniae. (A–H) ice (n = 5 per group) were infected with 2 × 105 CFU/mouse K. pneumoniae for 24 h and BALF was aseptically obtained. The cytokine profiles in BAL were assayed by ELISA. Data are shown as mean + SEM and are representative of three experiments. + p < 0.05, ++ p < 0.01 by one-way ANOVA.

Qiang Guo, et al. Eur J Immunol. 2012 June;42(6):1500-1511.
3.
Figure 4

Figure 4. From: Caveolin-1 plays a critical role in host immunity against Klebsiella pneumoniae by regulating STAT5 and Akt activity.

Cav1 deficiency altered the inflammatory response to K. pneumoniae at early times and in different organs. WT mice and cav1 KO mice (5 in each group) were infected with 2 × 105 CFU/mouse K. pneumoniae for (A–E) 8 h and (F–K) 24 h and the indicated organs were aseptically removed. (A–I) Cytokine/chemokine levels and (J and K) bacterial burden were evaluated. Data are shown as mean + SEM (n = 5 per group) and are representative of three experiments, + p < 0.01, ++ p < 0.001, one-way ANOVA.

Qiang Guo, et al. Eur J Immunol. 2012 June;42(6):1500-1511.
4.
Figure 2

Figure 2. From: Caveolin-1 plays a critical role in host immunity against Klebsiella pneumoniae by regulating STAT5 and Akt activity.

Severe lung injury in cav1 KO mice. WT and cav1 KO mice (n = 5 per group) were infected with 2 × 105 CFU/mouse K. pneumoniae for 24 h. (A) BALF was isolated and the percentage of neutrophils in total nucleated cells determined. (B) The lungs were fixed in formalin and sections were analyzed by H&E staining. One representative image of three experiments is shown. The arrows show regions of inflammation. (C) MPO levels were assessed in lung homogenates by HTAB. (D) ROS levels were assessed in lung homogenates by an H2DCF method. (A, C, and E) Data are shown as mean + SEM (n = 5 per group). + p = 0.044, ++ p = 0.02 by one-way ANOVA.

Qiang Guo, et al. Eur J Immunol. 2012 June;42(6):1500-1511.
5.
Figure 6

Figure 6. From: Caveolin-1 plays a critical role in host immunity against Klebsiella pneumoniae by regulating STAT5 and Akt activity.

MLE-12 cells expressing a dominant negative mutant Cav1 displayed an activated STAT5 pathway following K. pneumoniae infection. MLE-12 cells were either transfected with cav1 DN or cav1 WT plasmids or treated with 2 μM STAT5 inhibitor WC1066. After 24 h, the cells were infected with K. pneumoniae at 10:1 MOI for 1 h and surface bacteria were killed by incubating with polymyxin B for 1 h or left uninfected. (A) Bacterial burden determined as CFUs. (B) ROS levels were determined by an H2DCF assay. (C) Expression of the indicated molecules was determined by western blotting. GAPDH serves as the loading control. (D) Densitometry quantification of western blotting gel data presented in (C) using Quantity one software (one-way ANOVA (Tukey’s post-hoc), ++ p < 0.001). Data are (E) representative or shown (A, B, and D) as mean ± SEM of replicates of three representative experiments.

Qiang Guo, et al. Eur J Immunol. 2012 June;42(6):1500-1511.
6.
Figure 1

Figure 1. From: Caveolin-1 plays a critical role in host immunity against Klebsiella pneumoniae by regulating STAT5 and Akt activity.

Decreased survival rates and increased bacterial burdens in cav1 KO mice following K. pneumoniae infection. Cav1 KO mice (n = 6) and WT mice (n = 5) were intranasally infected with 2 × 105 CFU/mouse of K. pneumonia. (A) Survival was determined over time. Survival is represented by Kaplan–Meier survival curves (p = 0.029, 95% confidence interval: 11.7– 36.3, log-rank test). (B–E) Moribund cav1 KO mice (6 mice) and WT mice (5 mice) were euthanized for assessment of histological and biochemical alterations. (B and D) Lungs were aseptically removed and homogenized in PBS for analysis of bacterial burden at either 8 or 24-h postinfection. The same quantity of tissue was evaluated and the data are expressed as CFU/g tissue. (C and E) AMs were derived from BALF and lysed in PBS at either 8 or 24-h postinfection. Equal numbers of AMs were used to assess bacterial burdens. Data are shown as mean + SEM (n = 5 per group) and are representative of three experiments. ++ p < 0.001 by one-way ANOVA.

Qiang Guo, et al. Eur J Immunol. 2012 June;42(6):1500-1511.
7.
Figure 5

Figure 5. From: Caveolin-1 plays a critical role in host immunity against Klebsiella pneumoniae by regulating STAT5 and Akt activity.

Altered cellular signaling pathways in the lung with Cav1 deficiency. Mice were infected with 2 × 105 control CFU/mouse K. pneumoniae or sham (n = 5). (A and B) Lung tissues were lysed and STAT5, Akt, β-catenin, GSK3β, IL12a, and phospho-ERK1/2 levels determined by western blotting 24 h after infection. Representative blots of three experiments are shown. GAPDH serves as the loading control. (C) Densitometry quantification of the western blotting gel data presented in (A and B) using Quantity one software. Data are shown as mean + SEM; one-way ANOVA, (n = 5 per group) and are representative of three experiments + p < 0.05, ++ p < 0.01. (D and E) Lung tissues were lysed and the levels of the indicated molecules determined by western blotting 8 h postinfection (n = 5) by western blot analysis. (F) Densitometry quantification of the western blotting gel data presented in (D and E). (G) Immunohistochemistry with specific antibodies of lung tissue of cav1 KO mice and WT mice (n = 5 per group) before and after infection with K. pneumoniae for 24 h. Arrows indicate significant changes in fluorescent intensity between control and KO mice lungs. Data are representative of three experiments.

Qiang Guo, et al. Eur J Immunol. 2012 June;42(6):1500-1511.

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