Characterization of HeLa LoxP cells expressing doxycycline-inducible RFP-tagged constructs. (A) HeLa LoxP cells expressing various RFP-tagged dox-inducible constructs cells were untreated (−Dox) or treated with 2 μg/ml doxycycline for 48 h (+Dox). RFP was visualized by fluorescent microscopy to test for dox-dependent expression. The white dashed square represents the region of the image used for the inset. Scale bars: 20 μm. Inset scale bars: 3 μm. Scale bars: 5 μm. (B–D) HeLa LoxP cells expressing various RFP-tagged dox-inducible constructs cells were untreated (−Dox) or treated with 2 μg/ml doxycycline for 48 h (+Dox). The construct expression levels were then determined by RT-qPCR using CHMP4B-specific primers (single qPCR run with three technical replicates). (E) HeLa LoxP cells expressing various RFP-tagged dox-inducible constructs cells were treated with 2 μg/ml doxycycline for 48 h. The levels of expression among various constructs were the n compared using RT-qPCR with RFP specific primers (single qPCR run with three technical replicates). (F and G) HeLa LoxP cells expressing various RFP-tagged dox-inducible constructs cells were untreated (−Dox) or treated with 2 μg/ml doxycycline for 48 h (+Dox). The levels of RFP-tagged protein expressions were visualized via Western blot using anti-RFP and anti-Actin (loading control) antibodies. G shows Western blot quantification from three independent experiments. Statistical analysis is represented with a P value. Source data are available for this figure: SourceData FS6.