Cell culture dishes, plates, centrifuge tubes, and other plastic ware were purchased from BD Biosciences (Lincoln Park, NJ). Dulbecco’s modified Eagle’s medium (DMEM), Ham’s F-12, amphotericin B, gentamicin, and 0.25% trypsin/0.03% EDTA solution were from Invitrogen-Gibco (Grand Island, NY). Fetal bovine serum (FBS) was from Hyclone (Logan, UT). Bovine insulin, hydrocortisone, human EGF, cholera toxin A subunit, human transferrin, sodium selenite, dimethyl sulfoxide (DMSO) and Triton X-100 were from Sigma-Aldrich (St. Louis, MO). Mono-dansyl cadaverine (MDC) was from Invitrogen (Eugene, OR). OCT compound was obtained from Sakura Finetek USA. Inc. (Torrance, CA and the mounting medium (Immu-Mount) was from Thermo-Shandon (Pittsburgh, PA) Optisole-GS was purchased from Bausch and Lomb Inc. (Rochester, NY). Anti-fade medium (Gel/Mount M01) was from Biomeda Corp. (Foster City, CA).
The extraction reagent (TRIzol) was from Invitrogen-Gibco (Grand Island, NY). PCR buffer II (E12848) was obtained from Applied Biosystems (Foster City, CA). M-MuLV reverse transcriptase (Ready-To-Go You-Prime First-Strand Beads) were obtained from GE Healthcare, Inc. (Piscataway, NJ). PCR master mix (AmpErase UNG, Taqman MGB) probes for SPRR1A, SPRR1B, SPRR2A, and SPRR2B with assay IDs Hs00954595_s1, Hs00824893_m1, Hs01655189_sH, and Hs00863465_m1, respectively, were from Applied Biosystems.
The protein extraction kit (MPER) and BCA protein assay were from Pierce Biotechnology (Rockford, IL). Gel for protein electrophoresis (Ready Gel; 4%–15% Tris-HCl) was from Bio-Rad (Hercules, CA). Polyvinylidene difluoride (PVDF) membranes (Immobilon-P) were from Millipore (Billerica, MA). The electrophoretic transfer buffer was a Tris/glycine buffer from Bio-Rad. The TTBS wash buffer composed of 0.1% Tween 20 (Bio-Rad), 50 mM Tris and 0.15 M sodium chloride (pH 7.5). The chemiluminescent reagent (Luminol) was from Santa Cruz Biotechnology (Santa Cruz, CA).
Goat anti-rabbit AlexaFluor 488 antibodies (A11008) and goat anti-mouse AlexaFluor antibodies (A21151) were from Invitrogen-Molecular Probes (Carlsbad, CA). Goat anti-rabbit (no. 31460) and goat anti-mouse (no. 31430) horseradish peroxidase (HRP)–conjugated antibodies were from Pierce Biotechnology.