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After six TBS washes of 5 min each, sections were incubated overnight in different antisera.
A background correction was applied using the signal intensity resulting from Hoechst staining for each sections quantified.
Following three washing steps of 5 minutes each, sections were embedded in Vectashield mounting media containing DAPI.
After washing in PBS three times (3 min each), sections were rinsed with 0.3% Triton X-100 for 10 min followed by washing in PBS.
After washing in PBS thrice (3 min for each), sections were rinsed with 0.3% Triton X-100 for 10 min followed by washing in PBS.
After washing in PBS thrice (3 min for each), sections were treated with 0.04% DAB and 0.03%H2O22 at room temperature for visualization for 8 12 min.
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