- PA5-23065 - Invitrogen Antibodies KDM4C antibody

Submitted references RNA-seq analysis of PHD and VHL inhibitors reveals differences and similarities to the hypoxia response.

Frost J, Ciulli A, Rocha S
Wellcome open research 2019;4:17

Heterochromatin Reduction Correlates with the Increase of the KDM4B and KDM6A Demethylases and the Expression of Pericentromeric DNA during the Acquisition of a Transformed Phenotype.

Gurrion C, Uriostegui M, Zurita M
Journal of Cancer 2017;8(14):2866-2875

PITX1, a specificity determinant in the HIF-1α-mediated transcriptional response to hypoxia.

Mudie S, Bandarra D, Batie M, Biddlestone J, Moniz S, Ortmann B, Shmakova A, Rocha S
Cell cycle (Georgetown, Tex.) 2014;13(24):3878-91

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of JMJD2C using a polyclonal antibody (Product # PA5-23065).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of JMJD2C in HeLa nuclear extracts. Sample was incubated in JMJD2C polyclonal antibody (Product # PA5-23065).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of JMJD2C in 1.0 mg/mL HeLa lysate. Samples were incubated in JMJD2C polyclonal antibody (Product # PA5-23065). This experiment was performed under reducing conditions using the 12-230 kDa separation system.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of JMJD2C using a polyclonal antibody (Product # PA5-23065).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemical analysis of JMJD2C in paraffin embedded mouse pancreas. Samples were incubated in JMJD2C polyclonal antibody (Product # PA5-23065) followed by DAB with hematoxylin counterstain.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 2. PITX1 is a specificity determinant for HIF-1a-dependent target gene activation. (A) U2OS and HeLa cells were transfected with control or PITX1 siRNA, prior to treatment with 1% O2 for 24 hours. Whole cell lysates were obtained 48 hours post-transfection and analyzed by protein gel blot using the indicated antibodies. (B) HeLa cells were transfected with control or PITX1 siRNA, prior to treatment with 1% O2 for the indicated periods of time. Whole cell lysates were obtained 48 hours post-transfection and analyzed by western blot using the indicated antibodies. (C) HeLa cells were transfected, processed and analyzed as in B. (D) U2OS were co-transfected with 1 mug of GFP-HIF-1beta and increasing amounts of PITX1 plasmid (0.1, 0.25 and 0.5 mug) prior to exposure to 1% O2 for 24 hours. Whole cell lysates were obtained 48 hours post-transfection and analyzed by protein gel blot using the indicated antibodies. (E) MDA-MB-231-HRE cells were transfected with control and PITX1 siRNA oligonucleotides prior to treatment with 1% O2 for 24 hours. Luciferase activity was measured 48 hours post-transfection. Graph depicts mean and standard deviation of a minimum of 3 independent experiments. Student's t-test was performed to calculate p values, and levels of significance are denoted as follows: * P < 0.05, ** P < 0.01, and *** P < 0.001. (F) MDA-MB-231 cells were transfected with control and PITX1 siRNA oligonucleotides prior to treatment to 1% O2 for 24 hours. Whole cell lys

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 3. PITX1 regulates JMJD2B and JMJD2C expression in a HIF-1alpha-dependent manner. (A) HeLa cells were transfected with control or PITX1 siRNA, prior to treatment with 1% O2 for 24 hours. Total mRNA was extracted and levels of JMJD2A, JMJD2B and JMJD2C mRNA were analyzed by qPCR. Graph depicts mean and standard deviation of a minimum of 3 independent experiments. Data was normalized using actin and compared to control siRNA. Student's t-test was performed to calculate p values, and levels of significance are denoted as follows: * P < 0.05, ** P < 0.01, and *** P < 0.001. (B) HeLa cells were transfected with control or HIF-1alpha siRNA, prior to treatment with 1% O2 for 24 hours. Whole cell lysates were obtained 48 hours post-transfection and analyzed by protein gel blot using the indicated antibodies. (C) HeLa cells were transfected with control, PITX1, HIF-1a or PITX1 and HIF-1alpha siRNA, prior to treatment with 1% O2 for 24 hours. Whole cell lysates were obtained 48 hours post-transfection and analyzed by western blot using the indicated antibodies. (D) HeLa cells were transfected with the indicated siRNAs oligonucleotides prior to treatment with 1% O2 for 24 hours. Whole cell lysates were obtained 48 hours post-transfection and analyzed by protein gel blot using the indicated antibodies.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 3 Variation in the levels of enzymes that modify histone H3 in transformed MCF10A-Er-Src cell line . A, Immunostaining to detect KDM4B demethylase and western blots detecting KDM4A, KDM4B and KDM4C histone demethylases as well as the methyl transferase SUV39H1 levels in non-transformed and transformed cells. B, Immunostaining to detect UTX/KDM6A and western blot experiments detecting UTX/KDM6A and KDM6B in non-transformed and transformed cells. C, Immunostaining to detect the methyl transferase EZH2 and western blots to detect EZH2 and SUZ12 in transformed and no-transformed cells. Acting was used as loading control in all the western blot experiments. NT=Non-transformed cells; T= Transformed cells.

PA5-23065

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