Combination PPAR and RXR Agonist Treatment in Melanoma Cells: Functional Importance of S100A2
Figure 4
shS100A2 decreases S100A2 mRNA and protein in A375(DRO) cells. qRT-PCR and Western blot of S100A2. (a) One microgram of total RNA was used for the S100A2 quantitative reverse transcription-PCR analysis (ABIPRISM7700; Perkin-Elmer), and absolute values were derived from a standard curve using a known amount of sense strand RNA (ag, attograms of sense strand RNA). Isoform RNA was normalized to total input RNA (18s rRNA measured from 1 ng of total RNA). (b) 60 μg of nuclear protein extract from A375(DRO), the SCR shRNA infected control cell, and shS100A2 infected cells. Proteins were size-separated on a 10% SDS-PAGE gel and transferred to nitrocellulose. The blot was blocked with 10% nonfat milk and incubated with S100A2 primary antibody (sc Y20) and then secondary antibody with antimouse IgG conjugated to horse-radish peroxidase as previously described. β-actin was measured as a loading control.
