Expression of adrenergic receptors in HOK. (A) mRNA was isolated from HOK strains derived from three donors and hybridized to an Affymetrix HG-U133A microarray. The values in the table are signal intensities for hybridization with the microarray; values higher than 40 represent significant gene expression. Bold typeface indicates the 2 genes in the list that were significantly expressed. (B) Protein was isolated from HOK strains derived from four donors, and expression of the α2B-AR and β2-AR proteins was detected on a Western blot. Blots were stained with anti-α2B-AR (Genex Bioscience, Hayward, CA, USA) at a 1:5000 dilution (2.3 µg/mL), followed by HRP-linked anti-rabbit secondary antibody (Cell Signaling Technology, Danvers, MA, USA) at a 1:1000 dilution (0.2 µg/mL), or with anti-β2-AR (Abcam, Cambridge, MA, USA; clone Ab40834) at a 1:2500 dilution (0.2 µg/mL), followed by HRP-linked anti-goat secondary antibody (Abcam) at a 1:5000 dilution (0.4 µg/mL). α2B-AR was detected at 49 kDa as expected based on its amino acid composition. Immuno-detected β2-AR bands were evident at 49 kDa and 47 kDa, likely representing the palmitoylated and non-palmitoylated forms. Human epidermal keratinocytes were used as a positive control. 4, HOK from Donor 4; 5, HOK from Donor 5; 6, HOK from Donor 6; 7, HOK from Donor 7; 8, human epidermal keratinocytes from Donor 8.