Notebook Flow Cytometry

9-24-2009

• We performed standard measurements in HeLa, U2OS and MCF7 with our three standard promoters: CMV, JeT and JeT-CMV. These measurements will be performed in 3 independent measurements. The initial result of U2OS is shown below, which includes the following samples (see table 1).

Sample negative control (transfection with a non-fluorescent plasmid p6) cotransfection with JeT plasmid and p6 cotransfection with JeT_CMV plasmid and p6 cotransfection with CMV plasmid and p6 cotransfection with mCherry reference plasmid and p6 cotransfection with CMV plasmid and mCherry reference plasmid cotransfection with JeT plasmid and mCherry reference plasmid cotransfection with JeT_CMV plasmid and mCherry reference plasmid

Figure ?: Flow cytometry results of the U2-OS Standard plate (1st measurement). The Relative Expression Units (REU) are plotted against the different promoter/plasmid constructs. The three standard promoters are CMV, JeT, and JeT_CMV and they were measured 20 hours after transfection of the U2-OS cells.

• We measured the constitutive promoters in the HeLa cell line. Unfortunately, the number of the viable cells are not enough for a reliable measurement.

9-25-2009

• We measured the MCF-7 Standard plate, which includes the same samples as above (see table 1)

Figure ?: Flow cytometry results of the MCF-7 Standard plate (1sr measurement). The .

9-28-2009

• We measured the MCF-7 and the U2-OS Standard plate again (same samples).

Figure ?: Flow cytometry results of the U2-OS standard plate The .

Figure ?: Flow cytometry results of the MCF-7 standard plate The .

9-29-2009

• We measured the MCF-7 and the U2-OS Standard plate the third time (same samples).

Figure ?: Flow cytometry results of the U2-OS Standard plate (2nd measurement). The .

Figure ?: Flow cytometry results of the MCF-7 Standard plate (2nd measurement). The .

9-30-2009

• We measured the constitutive promoters in the HeLa cell line again.

Figure ?: Flow cytometry results of the constitutive promoters in HeLa (1st measurement). The .

10-02-2009

• We measured the NFkB responsive promoter plate in U2-OS 10 hours after induction.

Figure ?: Flow cytometry results of the NFkB responsive promoters in U2-OS (1st measurement). The .

10-03-2009

• We measured the HIF responsive promoter in MCF-7. Unfortunately, the cell number was too low.

10-05-2009

• We measured the HeLa Standard plate in three different experiments????, which includes the same samples as above (see table 1)
• We measured the constitutive promoters in the HeLa cell line again. Unfortunately, the number of the viable cells are not enough for a reliable measurement.
• We measured the AhR responsive promoters and the natural promoter CYP1A1 (AhR responsive promoter) in HeLa. Unfortunately, the number of the viable cells are not enough for a reliable measurement.

Figure ?: Flow cytometry results of the HeLa Standard plate (1st measurement). The .

Figure ?: Flow cytometry results of the HeLa Standard plate (2nd measurement). The .

Figure ?: Flow cytometry results of the HeLa Standard plate (3rd measurement). The .

10-07-2009

• We measured the NFkB responsive promoter plate in U2-OS 10 hours after induction again.

Figure ?: Flow cytometry results of the NFkB responsive promoters in U2-OS (2nd measurement). The .

10-08-2009

• We measured the Estrogen responsive promoter in MCF-7. Unfortunately, the number of the viable cells are not enough for a reliable measurement.

10-09-2009

• We measured the HeLa Standard plate (fourth time).
• We measured the SREBP responsive promoter in HeLa.
• We measured the constitutive promoters in HeLa again.

Figure ?: Flow cytometry results of the HeLa Standard plate (4th measurement). The .

File:.png Figure ?: Flow cytometry results of the SREBP responsive promoters in HeLa (1st measurement). The .

Figure ?: Flow cytometry results of the constitutive promoters in HeLa (2nd measurement).

10-11-2009

• We measured the natural promoter c-Jun (AP-1) in HeLa.

Figure ?: Flow cytometry results of the natural promoter c-Jun (AP-1). The .

10-12-2009

• We measured the HeLa Standard plate (fifth time).
• We measured the promosing NKkB responsive promoter NIIL10 in different medium in U2-OS. Here, we want to see, if there is an induction of a nonspecific drug (Thiazolidin??) and how the different medium affect the promoter activity. No significant change.

Figure ?: Flow cytometry results of the HeLa Standard plate (5th measurement). The .

Figure ?: Flow cytometry results of the NIIL10 promoter (NFkB responsive promoter) under different conditions in U2-OS. The .

10-13-2009

• We measured the p53 responsive promoter in MCF-7. Unfortunately, there were some problems with the flow cytometry analysis.

10-14-2009

• We measured