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Figure 4 | BMC Immunology

Figure 4

From: Novel Method of Monitoring Trace Cytokines and Activated STAT Molecules in the Paws of Arthritic Mice using Multiplex Bead Technology

Figure 4

Using Luminex® Multiplex Bead Arrays to Measure Intracellular PD Markers is More Sensitive than Conventional Western Blot Analysis. Using the 60 day CIA mouse paws we extracted the paw supernatants as described in the materials and methods and tested for phospho-STAT3 using either conventional western blot analysis with densitometry or multiplex bead analysis. (A) Extracts from two different mice per group were run on a SDS-PAGE gel and transferred onto a nitrocellulose membrane and exposed to film for the final image. Western blot was probed with anti-pSTAT3 α (79 kDa) and β (86 kDa) subunits, stripped then re-probed for total STAT3, α (76 kDα) and β (86 kDa) subunits as shown. Below graph shows densitometry analysis from above blot **ns p = 0.2037 versus vehicle, *ns p = 0.1226 versus CIA + vehicle group, N = 3 per group shown. (B) Luminex® phospho-STAT kit analyzed paw extractions from CIA mice treated with vehicle alone or dexamethasone. Samples were those same as used for the W.blot above, *p = 0.034 compared CIA + vehicle group, **p = 0.017 compared to vehicle alone. For both graphs, statistical test used for p-values: two-tailed paired Student's t-test, N = 3-4 mice per group shown. All graphs show Mean ± SEM, ns = not significant.

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