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Triple inducer systems. ( A ) Diagram of plasmids integrated into the CuRO-eGFP cell line. Cumate repressor (CymR), cumate operator (CuO), neomycin phosphotransferase II (G418), puromycin-N-acetyl-transferase (PAC), tetracycline repressor <t>(TetR),</t> T7 RNA polymerase (T7RNAP), vanillic acid repressor (VanR). ( B ) Western blot detection of eGFP and EF1α1 loading control from CuRO-eGFP clonal cell lines. A total of 2 × 10 6 cell equivalents were loaded per lane. ( C ) Diagram of plasmids integrated into the eGFP PHITER cell line. Annotation as described in panel A . ( D ) Western blot detection of eGFP and loading control EF1α1 following a 48 h induction of eGFP PHITER clones with 25 μg/mL Cym. A total of 2 × 10 6 cell equivalents were loaded per lane. ( E ) Diagram of plasmids integrated into the IBComp-eGFP PHIT cell line. Blasticidin S-deaminase (BSD), vanillic acid operator (VanO), tetracycline operators (TetO), bleomycin resistance gene (BLE). Other annotation as described in panel A. ( F ) Northern blot of total RNA from IBComp-eGFP PHIT clonal cell lines. Membrane was probed for individual repressor mRNAs, stripped, and then reprobed. tubulin , loading control.
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Triple inducer systems. ( A ) Diagram of plasmids integrated into the CuRO-eGFP cell line. Cumate repressor (CymR), cumate operator (CuO), neomycin phosphotransferase II (G418), puromycin-N-acetyl-transferase (PAC), tetracycline repressor <t>(TetR),</t> T7 RNA polymerase (T7RNAP), vanillic acid repressor (VanR). ( B ) Western blot detection of eGFP and EF1α1 loading control from CuRO-eGFP clonal cell lines. A total of 2 × 10 6 cell equivalents were loaded per lane. ( C ) Diagram of plasmids integrated into the eGFP PHITER cell line. Annotation as described in panel A . ( D ) Western blot detection of eGFP and loading control EF1α1 following a 48 h induction of eGFP PHITER clones with 25 μg/mL Cym. A total of 2 × 10 6 cell equivalents were loaded per lane. ( E ) Diagram of plasmids integrated into the IBComp-eGFP PHIT cell line. Blasticidin S-deaminase (BSD), vanillic acid operator (VanO), tetracycline operators (TetO), bleomycin resistance gene (BLE). Other annotation as described in panel A. ( F ) Northern blot of total RNA from IBComp-eGFP PHIT clonal cell lines. Membrane was probed for individual repressor mRNAs, stripped, and then reprobed. tubulin , loading control.
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Triple inducer systems. ( A ) Diagram of plasmids integrated into the CuRO-eGFP cell line. Cumate repressor (CymR), cumate operator (CuO), neomycin phosphotransferase II (G418), puromycin-N-acetyl-transferase (PAC), tetracycline repressor <t>(TetR),</t> T7 RNA polymerase (T7RNAP), vanillic acid repressor (VanR). ( B ) Western blot detection of eGFP and EF1α1 loading control from CuRO-eGFP clonal cell lines. A total of 2 × 10 6 cell equivalents were loaded per lane. ( C ) Diagram of plasmids integrated into the eGFP PHITER cell line. Annotation as described in panel A . ( D ) Western blot detection of eGFP and loading control EF1α1 following a 48 h induction of eGFP PHITER clones with 25 μg/mL Cym. A total of 2 × 10 6 cell equivalents were loaded per lane. ( E ) Diagram of plasmids integrated into the IBComp-eGFP PHIT cell line. Blasticidin S-deaminase (BSD), vanillic acid operator (VanO), tetracycline operators (TetO), bleomycin resistance gene (BLE). Other annotation as described in panel A. ( F ) Northern blot of total RNA from IBComp-eGFP PHIT clonal cell lines. Membrane was probed for individual repressor mRNAs, stripped, and then reprobed. tubulin , loading control.
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Triple inducer systems. ( A ) Diagram of plasmids integrated into the CuRO-eGFP cell line. Cumate repressor (CymR), cumate operator (CuO), neomycin phosphotransferase II (G418), puromycin-N-acetyl-transferase (PAC), tetracycline repressor <t>(TetR),</t> T7 RNA polymerase (T7RNAP), vanillic acid repressor (VanR). ( B ) Western blot detection of eGFP and EF1α1 loading control from CuRO-eGFP clonal cell lines. A total of 2 × 10 6 cell equivalents were loaded per lane. ( C ) Diagram of plasmids integrated into the eGFP PHITER cell line. Annotation as described in panel A . ( D ) Western blot detection of eGFP and loading control EF1α1 following a 48 h induction of eGFP PHITER clones with 25 μg/mL Cym. A total of 2 × 10 6 cell equivalents were loaded per lane. ( E ) Diagram of plasmids integrated into the IBComp-eGFP PHIT cell line. Blasticidin S-deaminase (BSD), vanillic acid operator (VanO), tetracycline operators (TetO), bleomycin resistance gene (BLE). Other annotation as described in panel A. ( F ) Northern blot of total RNA from IBComp-eGFP PHIT clonal cell lines. Membrane was probed for individual repressor mRNAs, stripped, and then reprobed. tubulin , loading control.
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Triple inducer systems. ( A ) Diagram of plasmids integrated into the CuRO-eGFP cell line. Cumate repressor (CymR), cumate operator (CuO), neomycin phosphotransferase II (G418), puromycin-N-acetyl-transferase (PAC), tetracycline repressor <t>(TetR),</t> T7 RNA polymerase (T7RNAP), vanillic acid repressor (VanR). ( B ) Western blot detection of eGFP and EF1α1 loading control from CuRO-eGFP clonal cell lines. A total of 2 × 10 6 cell equivalents were loaded per lane. ( C ) Diagram of plasmids integrated into the eGFP PHITER cell line. Annotation as described in panel A . ( D ) Western blot detection of eGFP and loading control EF1α1 following a 48 h induction of eGFP PHITER clones with 25 μg/mL Cym. A total of 2 × 10 6 cell equivalents were loaded per lane. ( E ) Diagram of plasmids integrated into the IBComp-eGFP PHIT cell line. Blasticidin S-deaminase (BSD), vanillic acid operator (VanO), tetracycline operators (TetO), bleomycin resistance gene (BLE). Other annotation as described in panel A. ( F ) Northern blot of total RNA from IBComp-eGFP PHIT clonal cell lines. Membrane was probed for individual repressor mRNAs, stripped, and then reprobed. tubulin , loading control.
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Triple inducer systems. ( A ) Diagram of plasmids integrated into the CuRO-eGFP cell line. Cumate repressor (CymR), cumate operator (CuO), neomycin phosphotransferase II (G418), puromycin-N-acetyl-transferase (PAC), tetracycline repressor <t>(TetR),</t> T7 RNA polymerase (T7RNAP), vanillic acid repressor (VanR). ( B ) Western blot detection of eGFP and EF1α1 loading control from CuRO-eGFP clonal cell lines. A total of 2 × 10 6 cell equivalents were loaded per lane. ( C ) Diagram of plasmids integrated into the eGFP PHITER cell line. Annotation as described in panel A . ( D ) Western blot detection of eGFP and loading control EF1α1 following a 48 h induction of eGFP PHITER clones with 25 μg/mL Cym. A total of 2 × 10 6 cell equivalents were loaded per lane. ( E ) Diagram of plasmids integrated into the IBComp-eGFP PHIT cell line. Blasticidin S-deaminase (BSD), vanillic acid operator (VanO), tetracycline operators (TetO), bleomycin resistance gene (BLE). Other annotation as described in panel A. ( F ) Northern blot of total RNA from IBComp-eGFP PHIT clonal cell lines. Membrane was probed for individual repressor mRNAs, stripped, and then reprobed. tubulin , loading control.
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Triple inducer systems. ( A ) Diagram of plasmids integrated into the CuRO-eGFP cell line. Cumate repressor (CymR), cumate operator (CuO), neomycin phosphotransferase II (G418), puromycin-N-acetyl-transferase (PAC), tetracycline repressor <t>(TetR),</t> T7 RNA polymerase (T7RNAP), vanillic acid repressor (VanR). ( B ) Western blot detection of eGFP and EF1α1 loading control from CuRO-eGFP clonal cell lines. A total of 2 × 10 6 cell equivalents were loaded per lane. ( C ) Diagram of plasmids integrated into the eGFP PHITER cell line. Annotation as described in panel A . ( D ) Western blot detection of eGFP and loading control EF1α1 following a 48 h induction of eGFP PHITER clones with 25 μg/mL Cym. A total of 2 × 10 6 cell equivalents were loaded per lane. ( E ) Diagram of plasmids integrated into the IBComp-eGFP PHIT cell line. Blasticidin S-deaminase (BSD), vanillic acid operator (VanO), tetracycline operators (TetO), bleomycin resistance gene (BLE). Other annotation as described in panel A. ( F ) Northern blot of total RNA from IBComp-eGFP PHIT clonal cell lines. Membrane was probed for individual repressor mRNAs, stripped, and then reprobed. tubulin , loading control.
Mouse Monoclonal Albumin Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Triple inducer systems. ( A ) Diagram of plasmids integrated into the CuRO-eGFP cell line. Cumate repressor (CymR), cumate operator (CuO), neomycin phosphotransferase II (G418), puromycin-N-acetyl-transferase (PAC), tetracycline repressor (TetR), T7 RNA polymerase (T7RNAP), vanillic acid repressor (VanR). ( B ) Western blot detection of eGFP and EF1α1 loading control from CuRO-eGFP clonal cell lines. A total of 2 × 10 6 cell equivalents were loaded per lane. ( C ) Diagram of plasmids integrated into the eGFP PHITER cell line. Annotation as described in panel A . ( D ) Western blot detection of eGFP and loading control EF1α1 following a 48 h induction of eGFP PHITER clones with 25 μg/mL Cym. A total of 2 × 10 6 cell equivalents were loaded per lane. ( E ) Diagram of plasmids integrated into the IBComp-eGFP PHIT cell line. Blasticidin S-deaminase (BSD), vanillic acid operator (VanO), tetracycline operators (TetO), bleomycin resistance gene (BLE). Other annotation as described in panel A. ( F ) Northern blot of total RNA from IBComp-eGFP PHIT clonal cell lines. Membrane was probed for individual repressor mRNAs, stripped, and then reprobed. tubulin , loading control.

Journal: mSphere

Article Title: A trypanosome trifecta: an independently tunable triple inducible system for genetic studies in Trypanosoma brucei

doi: 10.1128/msphere.00596-25

Figure Lengend Snippet: Triple inducer systems. ( A ) Diagram of plasmids integrated into the CuRO-eGFP cell line. Cumate repressor (CymR), cumate operator (CuO), neomycin phosphotransferase II (G418), puromycin-N-acetyl-transferase (PAC), tetracycline repressor (TetR), T7 RNA polymerase (T7RNAP), vanillic acid repressor (VanR). ( B ) Western blot detection of eGFP and EF1α1 loading control from CuRO-eGFP clonal cell lines. A total of 2 × 10 6 cell equivalents were loaded per lane. ( C ) Diagram of plasmids integrated into the eGFP PHITER cell line. Annotation as described in panel A . ( D ) Western blot detection of eGFP and loading control EF1α1 following a 48 h induction of eGFP PHITER clones with 25 μg/mL Cym. A total of 2 × 10 6 cell equivalents were loaded per lane. ( E ) Diagram of plasmids integrated into the IBComp-eGFP PHIT cell line. Blasticidin S-deaminase (BSD), vanillic acid operator (VanO), tetracycline operators (TetO), bleomycin resistance gene (BLE). Other annotation as described in panel A. ( F ) Northern blot of total RNA from IBComp-eGFP PHIT clonal cell lines. Membrane was probed for individual repressor mRNAs, stripped, and then reprobed. tubulin , loading control.

Article Snippet: TetR was detected using anti-TetR mouse monoclonal antibody (1:1,000, 1 h, Takara).

Techniques: Western Blot, Control, Clone Assay, Northern Blot, Membrane