Mast cell expression and HDAC expression on IgE exposure:
Mast cell were grown to confluence in six well cell culture plates exposed to 10ng/ml & 20 ng/ml concentration of IgE for 16 hours in serum free medium. Cells were then collected and centrifuged at 3000g, washed in 1X-PBS, lysed using RIPA buffer.  The lysate after protein estimation (Pierce™ BCA Protein Assay Kit Cat No:  23225, MA, USA) was used for western blot analysis (30 μg/μl) to assess endogenous Histone Deacetylase (HDAC) expression using Antibody Sampler Kit (#9928, Cell signalling Technology, MA, USA) using the corresponding antibodies namely, HDAC1, HDAC2, HDAC4, HDAC6 and 7 (representing Zinc dependent class I, class IIa and IIb ) as well as the Beta actin (β-Actin Antibody (AC-15) sc-69879, Santa Cruz Biotechnology, CA, USA) used as the loading control. Chemidoc-XRS+ (Biorad, CA, USA) was used for gel documentation and image lab tool (Biorad, CA, USA) was used for estimating the pixel density of the bands.