Fig. 6.

Chromosome mapping analysis of hGR cDNA. A, 10 μg of DNA from human placenta (lanes 1, 4), CHO/human somatic cell hybrid (HHW454, lanes 2, 5) containing chromosome 5 as its only human complement, or CHO (lanes 3, 6), was digested with EcoRI (lanes 1–3) or HindIII (lanes 4–6) to completion, fractionated on a 0.8% agarose gel and transferred to nitrocellulose paper. b, Chromosomes (3 × 10⁴) prepared from a human lymphocyte cell line, stained with 4,6-bis(2”-imadazolinyl-4H,5H)-2-phenylindole (DIPI)/chromomycin A3 and sorted using a dual-laser custom FACS IV chromosome sorter⁶³, were denatured and neutralized on nitrocellulose paper. Note that Hoechst/chromomycin-stained chromosome 9 was sorted with chromosomes 10–12. c, 10 μg of DNA from the parental mouse cell line MEL (lane 1) or the parentally derived somatic cell hybrid carrying human chromosome 16 (ref. 50; lane 2) was digested with HindIII and separated on a 0.8% agarose gel then transferred to nitrocellulose paper. All filters were probed with the 1,100-bp insert from hGR 1.2, nick-translated to a specific activity of 3 × 10⁸ c.p.m.μg^−l and hybridized in 5 × SSPE, 1 × Denhardt’s, 0.1% SDS, 50% formamide, 100 μg ml⁻¹ denatured salmon sperm DNA, 50% dextran sulphate at 42 °C for 18 h. Filters were washed twice (for 30 min each) in 2 × SSC at 68 °C and exposed to X-ray film at −70 °C with an intensifying screen.