Figure 2.

HDAC7 regulates CD36 transcription in endothelial cells. A, Nuclear HDAC7 translocation is increased in response to LPA exposure. MVECs were exposed to LPA (10 μM) for 24 hrs and then incubated with antibodies to HDAC5 or HADC7 followed by Alexa fluor® conjugated secondary antibodies. Cells were co-stained with DAPI to mark nuclei. In some studies cells were fractionated and the nuclear fractions isolated and subjected to Western blots for HDAC5 or HDAC7 expression. Representative results are shown from triplicate experiments. B., Knocking down endogenous HDAC7 attenuates LPA-mediated CD36 transcriptional repression in HMVECs. Western blots showed that HDAC 7 shRNA plasmids transfection effectively reduced the endogenous HDAC7 expression (upper panel). Cells transfected with HDAC7 shRNA plasmids or scrambled control were co-transfected with pGL3-CD36-luc and Renilla plasmids for 48 hrs. The cells were exposed to LPA (10 μM) or vehicle control for 24 hrs and luciferase activity measured as in Figure 1A (p < 0.01). C, HMVECs were transfected with a pool of HDAC7 shRNA plasmids for 28 hrs, and the cells were exposed to LPA (10 μM) for 18 hrs. The cells were harvested and total RNA was isolated for RT-PCR for CD36 mRNA expression. The image shows a representative gel from triplicate experiments.