Journal: International Journal of Molecular Sciences

Article Title: Disrupting the Btk Pathway Suppresses COPD-Like Lung Alterations in Atherosclerosis Prone ApoE −/− Mice Following Regular Exposure to Cigarette Smoke

doi: 10.3390/ijms19020343

Figure Lengend Snippet: Analysis of alveolar elastin breaks in lungs of ApoE −/− mice fed WD and exposed to SHS only (WD + SHS), with Btk inhibitor treatment (WD + SHS/Btk Inh.), or treated with endothelial cell targeted siRNA for MMP-9 (WD + SHS/MECA-32 siRNA MMP-9). Representative images are shown. Black arrows point out strand breaks in alveolar walls ( a ). Destruction of alveolar walls was assessed by counting numbers of breaks in elastin fibers ( b ). Groups receiving treatment scored significantly lower than the control group ( p < 0.05). Kruskal–Wallis ANOVA on ranks was used to assess statistical significance between groups ( p < 0.001) followed by post hoc testing with Dunn’s multiple comparison test for groups of interest, four to six mice per group were evaluated. * Significant difference detected.

Article Snippet: Treatment animals were injected intravenously [tail vein injection] with either Bruton’s tyrosine kinase inhibitor (BTK Inh) PCI-32765 (Selleck Chemicals, Houston, TX, USA) or siRNA specific for MMP-9 (Invitrogen, Carlsbad, CA, USA) conjugated with F(ab’) 2 fragments of anti-mouse neutrophil antibody (clone Ly-6G 1A8, Bio X Cell, West Lebanon, NH, USA) or anti-mouse endothelial cell antibody (clone MECA-32, Bio X Cell).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Disrupting the Btk Pathway Suppresses COPD-Like Lung Alterations in Atherosclerosis Prone ApoE −/− Mice Following Regular Exposure to Cigarette Smoke

doi: 10.3390/ijms19020343

Figure Lengend Snippet: ( a ) Morphometric analysis of lungs from ApoE −/− mice fed WD and exposed to SHS only (WD + SHS), with Btk inhibitor treatment (WD + SHS/Btk Inh.), or treated with endothelial cell targeted siRNA for MMP-9 (WD + SHS/MECA-32 siRNA MMP-9) showed reduced scores in treatment groups ( p < 0.05). Kruskal-Wallis ANOVA on ranks was used to assess statistical significance between groups ( p < 0.001) followed by post hoc testing with Dunn’s multiple comparison test for groups of interest, four to six animals per group were evaluated, with two groups of control animals combined. ( b ) Analysis of MLI in lungs of ApoE −/− mice fed WD and exposed to SHS or fed WD and exposed to SHS plus treatment with neutrophil targeted siRNA for MMP-9 (WD + SHS/Ly6G siRNA MMP-9) showed reduced scores in the treatment group ( p > 0.001). Student’s t -test was used to assess statistical significance, four mice per group were analyzed. * Significant difference detected.

Article Snippet: Treatment animals were injected intravenously [tail vein injection] with either Bruton’s tyrosine kinase inhibitor (BTK Inh) PCI-32765 (Selleck Chemicals, Houston, TX, USA) or siRNA specific for MMP-9 (Invitrogen, Carlsbad, CA, USA) conjugated with F(ab’) 2 fragments of anti-mouse neutrophil antibody (clone Ly-6G 1A8, Bio X Cell, West Lebanon, NH, USA) or anti-mouse endothelial cell antibody (clone MECA-32, Bio X Cell).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Disrupting the Btk Pathway Suppresses COPD-Like Lung Alterations in Atherosclerosis Prone ApoE −/− Mice Following Regular Exposure to Cigarette Smoke

doi: 10.3390/ijms19020343

Figure Lengend Snippet: Collagen deposition in lungs of ApoE −/− mice fed WD and exposed to SHS only (WD + SHS), with Btk inhibitor treatment (WD + SHS/Btk Inh.), or treated with endothelial cell targeted siRNA for MMP-9 (WD + SHS/MECA32 siRNA MMP-9). Picro-sirius red stained lung sections from ApoE −/− mice were viewed under regular light (left panels) or polarized light (right panels). Representative images are shown ( a ). Differences in hue of Picro-sirius red stained airways were evaluated using ImageJ software. Results are expressed as mean red component intensities (airway) or mean total intensity (vasculature) ( b ). Btk inhibition or EC targeted MMP-9 specific siRNA impacted collagen deposition in the airways or lung vasculature respectively. Kruskal-Wallis ANOVA on ranks was used to assess statistical significance in red intensity between groups ( p < 0.081). As Dunn’s multiple comparison test was not possible we assessed the reduction observed in the Btk inhibitor treated group using the non-parametric Mann-Whitney test with Bonferroni correction for multiple comparisons ( p = 0.019). For mean vascular intensity one way ANOVA was used to assess differences between groups ( p = 0.227) and as Fishers least significant difference test was not possible, we assessed the reduction observed in the MECA-32 siRNA MMP-9 treated group using Student’s t -test with Bonferroni correction for multiple comparison ( p = 0.097), four to six animals per group were evaluated, with two groups of control animals combined. * Significant difference detected.

Article Snippet: Treatment animals were injected intravenously [tail vein injection] with either Bruton’s tyrosine kinase inhibitor (BTK Inh) PCI-32765 (Selleck Chemicals, Houston, TX, USA) or siRNA specific for MMP-9 (Invitrogen, Carlsbad, CA, USA) conjugated with F(ab’) 2 fragments of anti-mouse neutrophil antibody (clone Ly-6G 1A8, Bio X Cell, West Lebanon, NH, USA) or anti-mouse endothelial cell antibody (clone MECA-32, Bio X Cell).

Techniques: Staining, Software, Inhibition, MANN-WHITNEY

Journal: International Journal of Molecular Sciences

Article Title: Disrupting the Btk Pathway Suppresses COPD-Like Lung Alterations in Atherosclerosis Prone ApoE −/− Mice Following Regular Exposure to Cigarette Smoke

doi: 10.3390/ijms19020343

Figure Lengend Snippet: Levels of MMP-9 in lung homogenates ( a ). Western blot from ApoE −/− mice fed WD and exposed to SHS only (WD + SHS), with Btk inhibitor treatment (WD + SHS/Btk Inh.), or treated with endothelial cell targeted siRNA for MMP-9 (WD + SHS/MECA-32 siRNA MMP-9). Three mice per group were analyzed ( b ). Western Blot from ApoE −/− mice fed WD and exposed to SHS only (WD + SHS) or treated with neutrophil targeted siRNA (WD + SHS/Ly6G siRNA MMP-9). Four animals per group were analyzed.

Article Snippet: Treatment animals were injected intravenously [tail vein injection] with either Bruton’s tyrosine kinase inhibitor (BTK Inh) PCI-32765 (Selleck Chemicals, Houston, TX, USA) or siRNA specific for MMP-9 (Invitrogen, Carlsbad, CA, USA) conjugated with F(ab’) 2 fragments of anti-mouse neutrophil antibody (clone Ly-6G 1A8, Bio X Cell, West Lebanon, NH, USA) or anti-mouse endothelial cell antibody (clone MECA-32, Bio X Cell).

Techniques: Western Blot