pparγ nbp2 22106 antibodies  (Novus Biologicals)

Journal: Cells

Article Title: RXR Agonists Enhance Lenalidomide Anti-Myeloma Activity and T Cell Functions while Retaining Glucose-Lowering Effect.

doi: 10.3390/cells12151993

Figure Lengend Snippet: Figure 3. LG100754 attenuates the binding effect of PPARα and PPAR γ on the CRBN promoter area. (A) U266 and MM1.R were transfected with CRBN/PGL3 firefly luciferase reported vector construct, then co-treated with PPARs agonist with LG100754 for 48 h, and luciferase bio-luminate activity was measured. (B) Bar graphs show qRT-PCR data using immunoprecipitated DNA obtained from ChIP with anti-CRBN or anti-IgG (negative control) antibodies; error bars represent SD. Results are presented as mean ± SD from at least three separate experiments. NS: not statistically significant; *: p < 0.05; **: p < 0.01.

Article Snippet: The PPARβ/δ (NBP2-22468) and PPARγ (NBP2-22106) antibodies were obtained from Novus Biologicals, LLC. (Littleton, CO, USA).

Techniques: Binding Assay, Transfection, Luciferase, Plasmid Preparation, Construct, Activity Assay, Quantitative RT-PCR, Immunoprecipitation, Negative Control

Journal: Cancer letters

Article Title: PPAR agonists attenuate lenalidomide's anti-myeloma activity in vitro and in vivo.

doi: 10.1016/j.canlet.2022.215832

Figure Lengend Snippet: Fig. 1. Treatment with PPAR agonists or overexpression of PPARs downregulated the mRNA and protein expression levels of CRBN. (A). Pathway enrichment analysis based on the microarray assay in MM cells treated with PIM kinase inhibitors; red arrows indicated PPAR pathway. (B). MM1.R, NCIH929 and RPMI8226 cells lines were treated with 5 μM or the EC50 concentration of individual PPAR agonist for 48 h, and levels of CRBN mRNA were measured by qRT-PCR. (C). MM1.R, NCIH929 and RPMI8226 cells were treated with 5 μM or the EC50 concentration of individual PPAR agonist for 48 h, and CRBN and GAPDH protein expression levels were assessed by Western blot. Bar graph represents the ratio of CRBN/GAPDH protein. One of 3 independent experiments was shown. (D). Representative images of DAPI and CRBN staining in NCIH929 cells treated with 5 μM of PPAR agonists for 48 h. (E). NCIH929 and MM1.R cells were transfected with a Lenti-PPARα, Lenti- PPARβ/δ, or Lenti-PPARγ expressing plasmid or control vector for 48 h. CRBN protein expression level was measured by Western blot.

Article Snippet: Slides were then incubated with anti-CD138 (1:20 dilution, Cat# ms-1793-3; Thermo Fisher, Waltham, MA), PPARα (1:400 dilution, Cat# SC-398394; Santa Cruz, Dallas, TX), PPARβ/δ (1:400 dilution, NBP2-22468; Novus, St. Louis, MO), or PPARγ (1:100 dilution, NBP2-22106; Novus) antibodies for 30 min at room temperature, followed by incubation with horseradish peroxidase (HRP)-labeled secondary antibody (Cat# K4001; DAKO, Santa Clara, CA).

Techniques: Over Expression, Expressing, Microarray, Concentration Assay, Quantitative RT-PCR, Western Blot, Staining, Transfection, Plasmid Preparation, Control

Journal: Cancer letters

Article Title: PPAR agonists attenuate lenalidomide's anti-myeloma activity in vitro and in vivo.

doi: 10.1016/j.canlet.2022.215832

Figure Lengend Snippet: Fig. 2. PPAR agonists reduce the anti-myeloma activity of lenalidomide in vitro. (A).

Article Snippet: Slides were then incubated with anti-CD138 (1:20 dilution, Cat# ms-1793-3; Thermo Fisher, Waltham, MA), PPARα (1:400 dilution, Cat# SC-398394; Santa Cruz, Dallas, TX), PPARβ/δ (1:400 dilution, NBP2-22468; Novus, St. Louis, MO), or PPARγ (1:100 dilution, NBP2-22106; Novus) antibodies for 30 min at room temperature, followed by incubation with horseradish peroxidase (HRP)-labeled secondary antibody (Cat# K4001; DAKO, Santa Clara, CA).

Techniques: Activity Assay, In Vitro

Journal: Cancer letters

Article Title: PPAR agonists attenuate lenalidomide's anti-myeloma activity in vitro and in vivo.

doi: 10.1016/j.canlet.2022.215832

Figure Lengend Snippet: Fig. 4. PPAR agonists reduce the anti-myeloma ac tivity of lenalidomide in vivo. (A). Bioluminescence intensity showing the different tumor burdens in NSG mice implanted with MM1.R myeloma cells and treated with control PBS (CTL), fenofibrate (Fen), lenalidomide (Len), or the combination (Len and Fen) (7 mice per group) from week 1 to week 10. (B). Left panel: the bioluminescence activity was quantified by determining the total flux (photons/sec) in each mouse from week 1 to week 10. Right panel: repre sentative images of tumors harvested from the control group (CTL) and the fenofibrate group (Fen) at week 8 and tumors harvested from the lenalidomide group (Len) and the combination group (Len and Fen) at week 10. (C). CRBN mRNA expression of the har vested tumors. qRT-PCR was used to detect the CRBN mRNA expression in the harvested tumors.

Article Snippet: Slides were then incubated with anti-CD138 (1:20 dilution, Cat# ms-1793-3; Thermo Fisher, Waltham, MA), PPARα (1:400 dilution, Cat# SC-398394; Santa Cruz, Dallas, TX), PPARβ/δ (1:400 dilution, NBP2-22468; Novus, St. Louis, MO), or PPARγ (1:100 dilution, NBP2-22106; Novus) antibodies for 30 min at room temperature, followed by incubation with horseradish peroxidase (HRP)-labeled secondary antibody (Cat# K4001; DAKO, Santa Clara, CA).

Techniques: In Vivo, Control, Activity Assay, Expressing, Quantitative RT-PCR

Journal: Cancer letters

Article Title: PPAR agonists attenuate lenalidomide's anti-myeloma activity in vitro and in vivo.

doi: 10.1016/j.canlet.2022.215832

Figure Lengend Snippet: Fig. 7. PPAR expression is upregulated in MM patients and is associated with poorer clinical outcomes. (A). Bone marrow biopsy sections from newly diagnosed MM patients and normal controls were stained with PPAR antibodies. Representative images of PPAR staining were shown. (B). Box plot shows the IHC scores of PPARs expression in CD138+ and CD138-cells in bone marrow biopsy samples of newly diagnosed myeloma patients and in normal control bone marrows. (C) Progression free survival (left) and overall survival (right) between newly diagnosed myeloma patients with high total PPAR IHC score (IHC score >8) and newly diagnosed myeloma patients with low total PPAR IHC score (IHC score ≤4). (D). Progression free survival and overall survival between patients with high PPARβ/δ expression (IHC >4) and patients with low PPARβ/δ expression (IHC ≤4). (E). Progression free survival and overall survival between patients with high PPARγ expression (IHC >4) and patients with low PPARγ expression (IHC ≤4). IHC scores represented the sum of score for CD138+ cells and score for CD138-cells.

Article Snippet: Slides were then incubated with anti-CD138 (1:20 dilution, Cat# ms-1793-3; Thermo Fisher, Waltham, MA), PPARα (1:400 dilution, Cat# SC-398394; Santa Cruz, Dallas, TX), PPARβ/δ (1:400 dilution, NBP2-22468; Novus, St. Louis, MO), or PPARγ (1:100 dilution, NBP2-22106; Novus) antibodies for 30 min at room temperature, followed by incubation with horseradish peroxidase (HRP)-labeled secondary antibody (Cat# K4001; DAKO, Santa Clara, CA).

Techniques: Expressing, Staining, Control