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. 2018 Aug 6;128(9):3887–3905. doi: 10.1172/JCI96393

Figure 1. An H2BeGFP pulse-chase system marks slow-cycling cells.

Figure 1

(A) Schematic representation of tetracycline-inducible lentiviral pSIN-TRE-H2BeGFP-rtTA2 construct. H2BeGFP is expressed via a tetracycline response element–containing promoter (TRE), which is activated by the reverse tetracycline transactivator (rtTA) induced in the presence of the tetracycline derivative doxycycline (DOX). RSV, constitutive promoter Rous sarcoma virus; RRE, Rev response element; cPPT, central polypurine tract; hPGK, human phosphoglycerate kinase promoter; WPRE, woodchuck hepatitis virus post-transcriptional regulatory element. (B) Chromatin accumulation of H2BeGFP in infected colon cancer cells upon DOX treatment. (C and D) Representative immunofluorescence picture of H2BeGFP-infected minitumors (MTs) growing embedded in Matrigel for colorectal (CRC) models or in suspension for melanoma (MEL) and glioblastoma (GBM) cancer models generated from single-cell suspensions. Cultures were treated with a DOX pulse chase to evaluate SCCCs together with cellular organization (phalloidin) (C) or proliferation (Ki67) (D). (C and D) Arrowheads, SCCCs. Hoechst was used as counterstain. (B–D) Scale bars: 100 μm.