Difference between revisions of "Part:BBa K2295002"
Line 6: Line 6:
  
  
−
[[Image:T-FREIBURG-HIF1A_shRNA2_134-50pc.png|900px|thumb|center|'''Figure 1:''' Flow cytometry of hypoxia response element promoter analysis. a)</b> Jurkat cells stably transduced with 4xCRE-pTal:eCFP were incubated 24 h in pH adjusted RPMI 1640. <b>b)</b> The experiment was repeated inducing with Forskolin (10 µM) and IBMX (10 µM). <b>c)</b> and <b>d)</b> HEK293T cells stably transduced with 4xCRE-pTal:eCFP with and without transient TDAG8 were induced similarly to <b>a)</b> and <b>b)</b>. Results show the represent the amount of eCFP positive cells <b>c)</b> and the <b>d)</b> mean fluorescence intensity. All data points are mean values of triplicates, error bars represent standard deviation. Significant differences in a) and b) were determined using ANOVA, for c) and d) significant differences were determined using one-tailed student’s t-test (Excel 2017); * p < 0.05, ** p < 0.01, *** p < 0.001, non-significant differences are not marked.]]
+
[[Image:T-FREIBURG-HIF1A_shRNA2_134-50pc.png|900px|thumb|center|'''Figure 1:''' <p><strong>Figure 7: Transient introduction of artificial HIF1A in HIF1A shRNA2 knockdown HEK293T cell line. </strong><br>
 +
                      Induction of the overexpressed HIF1A-HA can be observed in the last lane. Endogenous and artificial HIF1A were detected with HIF1A antibody; reintroduced HIF1A was detected via an HA-tag; GAPDH served as a loading control. Wild type (WT) and knockdown (HIF1A shRNA2) cells were transfected with CMV:HIF1A-HA one day prior to induction with CoCl<sub>2</sub> to simulate hypoxia. Cells were harvested 24 hours after induction. Immunoblot assay was performed using antibodies against HIF1A, HA-tag and GAPDH.</p>]]
  
  

Revision as of 03:06, 31 October 2017


HIF1A (Hypoxia-inducible factor one alpha)

Hif1a


Figure 1:

Figure 7: Transient introduction of artificial HIF1A in HIF1A shRNA2 knockdown HEK293T cell line.
Induction of the overexpressed HIF1A-HA can be observed in the last lane. Endogenous and artificial HIF1A were detected with HIF1A antibody; reintroduced HIF1A was detected via an HA-tag; GAPDH served as a loading control. Wild type (WT) and knockdown (HIF1A shRNA2) cells were transfected with CMV:HIF1A-HA one day prior to induction with CoCl2 to simulate hypoxia. Cells were harvested 24 hours after induction. Immunoblot assay was performed using antibodies against HIF1A, HA-tag and GAPDH.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 79
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 10
    Illegal AgeI site found at 1086
  • 1000
    COMPATIBLE WITH RFC[1000]