Human COUP-TF II/NR2F2 Antibody

Detection of Human COUP-TF II/NR2F2 by Knockdown Validated

Reduction of COUP-TFII or nucleolin decreases RAR beta2 transcription in MCF-7 cells.MCF-7 (A) and T47D (B) cells were transfected with control siRNA or an siRNA targeting nucleolin for 48 h. T47D cells were treated with EtOH or 1 µM atRA for 24 h. Q-PCR for nucleolin (NCL) and RARB2. Values are the average of triplicates. C, Western blot showing COUP-TFII and RAR beta2 expression after transfection with siCOUP-TFII. Values are relative to beta-actin. MCF-7 were transfected with siControl or siCOUP-TFII for 48 h and treated with 1 µM atRA for 6 h. Q-PCR was also performed for RRIG1. P<0.001 * versus control or ** versus atRA. Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0038278), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human COUP-TF II/NR2F2 by Knockdown Validated

Loss of miR-101 promotes cancer metastasis through de-repression of COUP-TFII expression.(a) LNCaP and PC3 cells were individually treated with 50 nM of miR-101 and miR-27a mimic for 48 h and then invasion assays were performed for an additional 16 h (n=3). *P<0.05 (two-sided Student's t-test) compared with the control group. (b) Representative western blot showed the levels of COUP-TFII, E-cadherin, vimentin and beta-actin in PC3 cells stably knocked down by COUP-TFII. (c) LNCaP and PC3 cells stably knocked-down of COUP-TFII were used to perform cell invasion assay. Invaded cells were stained and counted (lower panel) (n=3). Representative western blot showed the knockdown efficiency of COUP-TFII (upper panel). *: P<0.05 (two-sided Student's t-test) compared with control group. (d) PC3 cells carrying an inducible miR-101 in the absence and presence of doxycycline, and re-expression of COUP-TFII were used to perform invasion assays (n=3). Representative western blot showed the levels of COUP-TFII and beta-actin (left). Invaded cells were counted and results are shown in the right panel. *: P<0.05 (two-sided Student's t-test) (e) 22RV-1 cells were treated with miR-101 inhibitor (antisense RNA) in conjunction with knockdown of COUP-TFII and used for invasion assays (n=3). Representative western blot show the levels of COUP-TFII and beta-actin (left). Invaded cells were counted and result is shown in the right panel. *: P<0.05 (two-sided Student's t-test). (f) LNCaP cells containing a construct with inducible expression of COUP-TFII shRNA and constructs expressing with anti-miR-101 and anti-mir-27a were orthotopically injected into NOD-SCID mouse prostate. In addition these cells also contain a luciferase reporter to detect cancer cells. After the tumour size was bigger than 50 mm3, drinking water with or without doxycycline (1 mg ml−1) was given to the mice for 6 weeks to induce the COUP-TFII shRNA to repress COUP-TFII expression. Representative bioluminescence results show the status of tumour growth in different groups. (g) Quantification result of luminance from IVIS (control: n=5; anti-miR-101/27a(−): n=6; anti-miR-101/27a(+): n=7. *: P<0.05 (two-sided Student's t-test). (h) Immunohistochemical stain showed the metastatic tumour cells located in the mouse lymph node are positive for AR. Scale bar: 100 μM (low power); and 200 μM (high power). SCID, severe combined immunodeficient. Image collected and cropped by CiteAb from the following publication (https://www.nature.com/articles/ncomms11418), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human COUP-TF II/NR2F2 by Knockdown Validated

Loss of miR-101 promotes cancer metastasis through de-repression of COUP-TFII expression.(a) LNCaP and PC3 cells were individually treated with 50 nM of miR-101 and miR-27a mimic for 48 h and then invasion assays were performed for an additional 16 h (n=3). *P<0.05 (two-sided Student's t-test) compared with the control group. (b) Representative western blot showed the levels of COUP-TFII, E-cadherin, vimentin and beta-actin in PC3 cells stably knocked down by COUP-TFII. (c) LNCaP and PC3 cells stably knocked-down of COUP-TFII were used to perform cell invasion assay. Invaded cells were stained and counted (lower panel) (n=3). Representative western blot showed the knockdown efficiency of COUP-TFII (upper panel). *: P<0.05 (two-sided Student's t-test) compared with control group. (d) PC3 cells carrying an inducible miR-101 in the absence and presence of doxycycline, and re-expression of COUP-TFII were used to perform invasion assays (n=3). Representative western blot showed the levels of COUP-TFII and beta-actin (left). Invaded cells were counted and results are shown in the right panel. *: P<0.05 (two-sided Student's t-test) (e) 22RV-1 cells were treated with miR-101 inhibitor (antisense RNA) in conjunction with knockdown of COUP-TFII and used for invasion assays (n=3). Representative western blot show the levels of COUP-TFII and beta-actin (left). Invaded cells were counted and result is shown in the right panel. *: P<0.05 (two-sided Student's t-test). (f) LNCaP cells containing a construct with inducible expression of COUP-TFII shRNA and constructs expressing with anti-miR-101 and anti-mir-27a were orthotopically injected into NOD-SCID mouse prostate. In addition these cells also contain a luciferase reporter to detect cancer cells. After the tumour size was bigger than 50 mm3, drinking water with or without doxycycline (1 mg ml−1) was given to the mice for 6 weeks to induce the COUP-TFII shRNA to repress COUP-TFII expression. Representative bioluminescence results show the status of tumour growth in different groups. (g) Quantification result of luminance from IVIS (control: n=5; anti-miR-101/27a(−): n=6; anti-miR-101/27a(+): n=7. *: P<0.05 (two-sided Student's t-test). (h) Immunohistochemical stain showed the metastatic tumour cells located in the mouse lymph node are positive for AR. Scale bar: 100 μM (low power); and 200 μM (high power). SCID, severe combined immunodeficient. Image collected and cropped by CiteAb from the following publication (https://www.nature.com/articles/ncomms11418), licensed under a CC-BY license. Not internally tested by R&D Systems.