Measurement of changes in mRNA and protein expression of HDAC 6. Measurement of changes in mRNA and protein expression of HDAC 6 by SNC-121 treatment in normal and ocular hypertensive animals at day 7 (a,
c) and day 42 (b,
d) post-injury. Total RNA was extracted from retina using TRIzol reagent and 1 µg RNA was used to synthesize cDNA. The mRNA levels were examined by quantitative RT-PCR using primers specific to HDAC 6 and relative gene expression was calculated based on the comparative threshold cycle (Ct) method, normalized to housekeeping genes, β-actin a and b. Retina lysates (10 µg) were analyzed by Western blotting using anti-HDAC 6, and anti-β-actin antibodies. The band intensities were measured using chemiluminescent reagent and Versadoc imaging system c and d. Data expressed as mean ± SE. *P < 0.05; **P < 0.01; ****P < 0.0001; n = 3–21.