Journal: PLoS ONE
Article Title: Bcl-2 Inhibits the Innate Immune Response during Early Pathogenesis of Murine Congenital Muscular Dystrophy
doi: 10.1371/journal.pone.0022369
Figure Lengend Snippet: ( A–F , M–R ) Hindlimbs were immunostained with antibodies to TN-C ( A , C , E ) and LYVE-1 ( B , D , F ). ( A ) MyoD-hBcl-2 transgenic Lama2 +/− mice had strong staining of epimysium and tendons but negligible levels of TN-C around muscle fibers, similar to non-transgenic normal mice at this age. ( C ) TN-C expression was elevated around Lama2 −/− muscle fibers, but this was reduced in MyoD-hBcl-2 transgenic Lama2 −/− muscles ( E ). ( B ) Lama2 +/− ; MyoD-hBcl-2 +/− mice had a normal punctate pattern of LYVE-1 expression, marking lymphatic capillaries within the muscle bed and interstitium as in non-transgenic normal mice. ( D ) LYVE-1-expression was reduced in capillaries within the muscle bed of Lama2 −/− mice, and this was not affected by the MyoD-hBcl-2 transgene at 7 days of age ( F ). ( G and H ) Quantitation of TN-C and LYVE-1 expression in 7 day-old Lama2- expressing ( Lama2(+) : Lama2 +/+ or +/− ) muscle either with or without MyoD-hBcl-2 transgene showed there was no effect of transgene on these markers in normal muscle (n = 4–5, p>0.05). ( I and J ) Quantitation of TN-C and LYVE-1 expression in 7 day-old Lama2(+) and Lama2(−) (Lama2 −/− ) muscle with or without MyoD-hBcl-2 transgene (n = 7–9). ( K and L ) Quantitation of TN-C and LYVE-1 in 14 day-old muscles (n = 3–4) (* p<0.03, ** p<0.001). ( M–R ) Higher magnification images of muscle samples show that reduced LYVE-1 expression occurs more broadly throughout the muscle bed of Lama2 -deficient mice than elevated expression of TN-C. ( M , N ) MyoD-hBcl-2 transgenic Lama2 +/− muscle in panels 1A and 1B showing normal expression of TN-C and LYVE-1, respectively. ( P , Q ) MyoD-hBcl-2 transgenic Lama2 −/− muscle in panels 1E, and 1F, showing elevated TN-C and reduced LYVE-1 staining. ( O ) and ( R ) are merged images. (Scale bars in all images: 100 µM, T: tibia).
Article Snippet: Primary antibodies used were rat anti-mouse TN-C (diluted 1/500, clone MTn-12, T3413, Sigma, St. Louis, MO, USA,), rabbit anti-mouse LYVE-1 (1/500, 14917, Abcam, Cambridge, MA, USA), rat anti-mouse major basic protein (MBP) (1/1000, clone MT-14-7-3, from J.J. Lee, Mayo Clinic) and Alexa Fluor 488-labeled rat anti-mouse CD11b (1/500, M1/70, 557672, BD Pharmingen, San Diego, CA, USA).
Techniques: Transgenic Assay, Staining, Expressing, Quantitation Assay
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