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nr4a1 antibody  (Novus Biologicals)
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Novus Biologicals nr4a1 antibody
Figure 1. Induction of IL24 by <t>NR4A1</t> antagonist. A, Rh30 cells were treated with CDIM8 in the absence () or presence of IL24 knockdown (siIL-24), and whole-cell lysates were analyzed by Western blots. B and C, Rh30 cells were treated with 20 mmol/L CDIM8 for different times or for 24 hours (B) and whole-cell lysates were analyzed by Western blots (C). D, ARMS cells were transfected with pCMV-IL24 expression plasmid and after 24 hours, whole-cell lysates were analyzed by Western blots. E, Tumor lysates from athymic nude mice bearing Rh30 cells as xenografts (39) and treated with corn oil (control) or adenoviral – IL24 were analyzed by Western blots. Results are expressed as means SD for at three replicate determinations for each treatment group (E) and significant (P < 0.05) effects () are indicated.
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Figure 1. Induction of IL24 by NR4A1 antagonist. A, Rh30 cells were treated with CDIM8 in the absence () or presence of IL24 knockdown (siIL-24), and whole-cell lysates were analyzed by Western blots. B and C, Rh30 cells were treated with 20 mmol/L CDIM8 for different times or for 24 hours (B) and whole-cell lysates were analyzed by Western blots (C). D, ARMS cells were transfected with pCMV-IL24 expression plasmid and after 24 hours, whole-cell lysates were analyzed by Western blots. E, Tumor lysates from athymic nude mice bearing Rh30 cells as xenografts (39) and treated with corn oil (control) or adenoviral – IL24 were analyzed by Western blots. Results are expressed as means SD for at three replicate determinations for each treatment group (E) and significant (P < 0.05) effects () are indicated.

Journal: Molecular Cancer Research

Article Title: Inhibition of NR4A1 Promotes ROS Accumulation and IL24-Dependent Growth Arrest in Rhabdomyosarcoma

doi: 10.1158/1541-7786.mcr-19-0408

Figure Lengend Snippet: Figure 1. Induction of IL24 by NR4A1 antagonist. A, Rh30 cells were treated with CDIM8 in the absence () or presence of IL24 knockdown (siIL-24), and whole-cell lysates were analyzed by Western blots. B and C, Rh30 cells were treated with 20 mmol/L CDIM8 for different times or for 24 hours (B) and whole-cell lysates were analyzed by Western blots (C). D, ARMS cells were transfected with pCMV-IL24 expression plasmid and after 24 hours, whole-cell lysates were analyzed by Western blots. E, Tumor lysates from athymic nude mice bearing Rh30 cells as xenografts (39) and treated with corn oil (control) or adenoviral – IL24 were analyzed by Western blots. Results are expressed as means SD for at three replicate determinations for each treatment group (E) and significant (P < 0.05) effects () are indicated.

Article Snippet: NR4A1 antibody was purchased from Novus Biologicals.

Techniques: Knockdown, Western Blot, Transfection, Expressing, Plasmid Preparation, Control

Figure 2. Anticarcinogenic effects of NR4A1 inactivation are due to induction of IL24. Rh30 cells were transfected with siNR4A1 or treated with 20 mmol/L CDIM8 in the absence or presence of siIL24; effects on protein expression (A) cell proliferation (B), induction of Annexin V staining (C), and cell migration (D) were determined as outlined in the Materials and Methods section. Results are expressed as means SD for three replicate determinations for each treatment group (B, C þ D) and significant (P < 0.05) effects () and reversal of these responses () are indicated.

Journal: Molecular Cancer Research

Article Title: Inhibition of NR4A1 Promotes ROS Accumulation and IL24-Dependent Growth Arrest in Rhabdomyosarcoma

doi: 10.1158/1541-7786.mcr-19-0408

Figure Lengend Snippet: Figure 2. Anticarcinogenic effects of NR4A1 inactivation are due to induction of IL24. Rh30 cells were transfected with siNR4A1 or treated with 20 mmol/L CDIM8 in the absence or presence of siIL24; effects on protein expression (A) cell proliferation (B), induction of Annexin V staining (C), and cell migration (D) were determined as outlined in the Materials and Methods section. Results are expressed as means SD for three replicate determinations for each treatment group (B, C þ D) and significant (P < 0.05) effects () and reversal of these responses () are indicated.

Article Snippet: NR4A1 antibody was purchased from Novus Biologicals.

Techniques: Transfection, Expressing, Staining, Migration

Figure 3. Role of ROS in mediating the anticancer activities of NR4A1 silencing or CDIM8. A, Rh30 cells were transfected with siNR4A1 or treated with CDIM8 in the presence or absence of 5 mmol/L GSH and induction of ROS was determined by fluorescence using the cell- permeable probe as outlined in the Materials and Methods section. B, Cells were treated as described in A and effects on protein expression (from whole- cell lysates) were determined by Western blots. Rh30 cells were treated as described in A and effects on cell proliferation (C), Annexin V staining (D), and migration (E) were determined as outlined in the Materials and Methods section. Results are expressed as means SD for three replicate determinations for each treatment group (A, C, D þ E) and significant (P < 0.05) effects () and reversal of these responses () are indicated.

Journal: Molecular Cancer Research

Article Title: Inhibition of NR4A1 Promotes ROS Accumulation and IL24-Dependent Growth Arrest in Rhabdomyosarcoma

doi: 10.1158/1541-7786.mcr-19-0408

Figure Lengend Snippet: Figure 3. Role of ROS in mediating the anticancer activities of NR4A1 silencing or CDIM8. A, Rh30 cells were transfected with siNR4A1 or treated with CDIM8 in the presence or absence of 5 mmol/L GSH and induction of ROS was determined by fluorescence using the cell- permeable probe as outlined in the Materials and Methods section. B, Cells were treated as described in A and effects on protein expression (from whole- cell lysates) were determined by Western blots. Rh30 cells were treated as described in A and effects on cell proliferation (C), Annexin V staining (D), and migration (E) were determined as outlined in the Materials and Methods section. Results are expressed as means SD for three replicate determinations for each treatment group (A, C, D þ E) and significant (P < 0.05) effects () and reversal of these responses () are indicated.

Article Snippet: NR4A1 antibody was purchased from Novus Biologicals.

Techniques: Transfection, Expressing, Western Blot, Staining, Migration

Figure 4. Role of ROS and IL24 in mediating the anticancer activities of NR4A1 inactivation by siNR4A1 or CDIM8 in RD cells. A, RD cells were transfected with siNR4A1 or treated with CDIM8 in the presence or absence of 5 mmol/L GSH, and ROS was determined using the cell-permeable fluorescent probe as outlined in the Materials and Methods section. RD cells were treated as described in A and effects on protein expression (B), cell proliferation (C), Annexin V staining (D), and cell migration (E) were determined as outlined in the Materials and Methods section. F, RD cells were transfected with siNR4A1 or treated with CDIM8 in the presence or absence of siIL24 (cotransfected) and whole-cell lysates were analyzed by Western blots. RD cells were treated as outlined in F and effects on cell proliferation (G), Annexin V staining (H), and cell migration (I) were determined as outlined in the Materials and Methods section. Results are expressed as means SD for three replicate determinations for each treatment group (A, C, D, E, G þ H) and significant (P < 0.05) effects () and reversal of these responses () are indicated.

Journal: Molecular Cancer Research

Article Title: Inhibition of NR4A1 Promotes ROS Accumulation and IL24-Dependent Growth Arrest in Rhabdomyosarcoma

doi: 10.1158/1541-7786.mcr-19-0408

Figure Lengend Snippet: Figure 4. Role of ROS and IL24 in mediating the anticancer activities of NR4A1 inactivation by siNR4A1 or CDIM8 in RD cells. A, RD cells were transfected with siNR4A1 or treated with CDIM8 in the presence or absence of 5 mmol/L GSH, and ROS was determined using the cell-permeable fluorescent probe as outlined in the Materials and Methods section. RD cells were treated as described in A and effects on protein expression (B), cell proliferation (C), Annexin V staining (D), and cell migration (E) were determined as outlined in the Materials and Methods section. F, RD cells were transfected with siNR4A1 or treated with CDIM8 in the presence or absence of siIL24 (cotransfected) and whole-cell lysates were analyzed by Western blots. RD cells were treated as outlined in F and effects on cell proliferation (G), Annexin V staining (H), and cell migration (I) were determined as outlined in the Materials and Methods section. Results are expressed as means SD for three replicate determinations for each treatment group (A, C, D, E, G þ H) and significant (P < 0.05) effects () and reversal of these responses () are indicated.

Article Snippet: NR4A1 antibody was purchased from Novus Biologicals.

Techniques: Transfection, Expressing, Staining, Migration, Western Blot

Figure 5. Regulation of TXNDC5 and IDH1 by NR4A1 and the effects of knockdown of TXNDC5 and IDH1 on ROS. RD and Rh30 cells were transfected with siNR4A1 (A) or treated with 20 mmol/L CDIM8 (B) in the presence or absence of GSH and after 24 hours, whole-cell lysates were analyzed by Western blots. RD and Rh30 cells were transfected with siTXNDC5 (C) or siIDH-1 (D) to knock down the TXNDC5 and IDH-1 genes, respectively, and induction of ROS was determined using the fluorescent cell-permeable probe. Results are expressed as means SD for three replicate determinations for each treatment group (C þ D) and significant (P < 0.05) effects () and reversal of these responses () are indicated.

Journal: Molecular Cancer Research

Article Title: Inhibition of NR4A1 Promotes ROS Accumulation and IL24-Dependent Growth Arrest in Rhabdomyosarcoma

doi: 10.1158/1541-7786.mcr-19-0408

Figure Lengend Snippet: Figure 5. Regulation of TXNDC5 and IDH1 by NR4A1 and the effects of knockdown of TXNDC5 and IDH1 on ROS. RD and Rh30 cells were transfected with siNR4A1 (A) or treated with 20 mmol/L CDIM8 (B) in the presence or absence of GSH and after 24 hours, whole-cell lysates were analyzed by Western blots. RD and Rh30 cells were transfected with siTXNDC5 (C) or siIDH-1 (D) to knock down the TXNDC5 and IDH-1 genes, respectively, and induction of ROS was determined using the fluorescent cell-permeable probe. Results are expressed as means SD for three replicate determinations for each treatment group (C þ D) and significant (P < 0.05) effects () and reversal of these responses () are indicated.

Article Snippet: NR4A1 antibody was purchased from Novus Biologicals.

Techniques: Knockdown, Transfection, Western Blot

Figure 7. IL24-induced anticancer activities are due to silencing of TXNDC5 and IDH-1. RD and Rh30 cells were transfected with siTXNDC5 (A) or siIDH-1 (B) in the presence or absence of siIL24 and whole-cell lysates were analyzed by Western blots. RD and Rh30 cells were treated as described in A and B in the presence or absence of cotransfected siIL24 and the effects on cell proliferation (C), Annexin V staining (D), and cell migration (E) were determined as outlined in the Materials and Methods section. F, Model describing the anticancer activities of NR4A1 antagonists. Results are expressed as means SD for three replicate determinations for each treatment group (C, D þ E) and significant (P < 0.05) effects () and reversal of these responses () are indicated.

Journal: Molecular Cancer Research

Article Title: Inhibition of NR4A1 Promotes ROS Accumulation and IL24-Dependent Growth Arrest in Rhabdomyosarcoma

doi: 10.1158/1541-7786.mcr-19-0408

Figure Lengend Snippet: Figure 7. IL24-induced anticancer activities are due to silencing of TXNDC5 and IDH-1. RD and Rh30 cells were transfected with siTXNDC5 (A) or siIDH-1 (B) in the presence or absence of siIL24 and whole-cell lysates were analyzed by Western blots. RD and Rh30 cells were treated as described in A and B in the presence or absence of cotransfected siIL24 and the effects on cell proliferation (C), Annexin V staining (D), and cell migration (E) were determined as outlined in the Materials and Methods section. F, Model describing the anticancer activities of NR4A1 antagonists. Results are expressed as means SD for three replicate determinations for each treatment group (C, D þ E) and significant (P < 0.05) effects () and reversal of these responses () are indicated.

Article Snippet: NR4A1 antibody was purchased from Novus Biologicals.

Techniques: Transfection, Western Blot, Staining, Migration