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. Author manuscript; available in PMC: 2016 Feb 14.
Published in final edited form as: Nat Commun. 2015 Aug 14;6:8005. doi: 10.1038/ncomms9005

Figure 3.

Figure 3

Enhanced HDAC4 protein stability is correlated with greater rod protection efficiency. (a) Protein decay assay of HDAC4 Full Length and deletion alleles. HEK 293T cells were transfected with CAG-HDAC4 or CAG-HDAC4 deletion constructs for 20 hours before treatment with cycloheximide (100μg/ml) for 3 or 6 hours. Cell lysates were immunoblotted for HDAC4. The same immunoblot membranes were stripped and re-probed for actin. (b) Protein levels were quantified by densitometry after normalization to actin. Data are presented as mean ± s.d., n=3. (c) Protein decay assay for HDAC4 and HDAC4-S298A/S302A mutant. HEK293T cells were transfected with CAG-HDAC4 or CAG-HDAC4-S298A/S302A for 20 hours before treatment with cycloheximide (100μg/ml) for 3 or 6 hours. Cell lysates were immunoblotted for HDAC4. The same immunoblot membranes were stripped and re-probed for actin. (d) Protein levels were quantified by densitometry after normalization to actin. Data are presented as mean ± s.d., n=3. (e) HDAC4-S298A/S302A saved more rd1 rods in comparison to the wild type HDAC4 at P50. Scale bar, 40 μm. (f) Quantification of the preserved rods per 25600 μm2 on the flat-mount retina. *P<0.01. Data are presented as mean ± s.d., n=3.