mab177  (R&D Systems)

Journal: Cells

Article Title: Partial Inhibition of Epithelial-to-Mesenchymal Transition (EMT) Phenotypes by Placenta-Derived DBMSCs in Human Breast Cancer Cell Lines, In Vitro

doi: 10.3390/cells13242131

Figure Lengend Snippet: Modulation in chemokine and cytokine expression in DBMSCs after treatment with MDA231 cells.

Article Snippet: Fluorescent-labelled antibodies for flow cytometry experiments, including Human Snail Alexa Fluor ® 488-conjugated Antibody, Cat# IC3639G; Human/Mouse/Rat Vimentin APC-conjugated Antibody, Cat# IC2105A; Human Fibronectin PE-conjugated Antibody, Cat# IC1918P; Human/Mouse Wnt-5a Antibody (Cat# MAB645); Human IL-27 PE-conjugated Antibody, Cat# IC25261P; Human BAFF/BLyS/TNFSF13B PE-conjugated Antibody, Cat# IC1241P, Human IL-17RA/IL-17R Antibody; Cat# MAB177; Mouse F(ab)2 IgG (H+L) Fluorescein-conjugated Antibody, Cat# F0103B; Rabbit IgG Fluorescein-conjugated Antibody, Cat# F0112; and Goat IgG (H+L) Fluorescein-conjugated Antibody, Cat# F0109, were purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Expressing, Migration, Activity Assay

Journal: Cells

Article Title: Partial Inhibition of Epithelial-to-Mesenchymal Transition (EMT) Phenotypes by Placenta-Derived DBMSCs in Human Breast Cancer Cell Lines, In Vitro

doi: 10.3390/cells13242131

Figure Lengend Snippet: Effect of cancer cell treatment on the expression of chemokines and cytokines on DBMSCs: Flow cytometry analysis for expression of chemokines and cytokines by DBMSCs after co-culture with DBMSCs showed a significant increase in IL-21, IFNA2, TNSF13B, and IL-17R for 96 h ( A ) but a significant decrease in the expression level of IL-27. The mean fluorescence index (MFI) is plotted and shown as a bar diagram ( B ). Bars represent standard errors. * p < 0.05.

Article Snippet: Fluorescent-labelled antibodies for flow cytometry experiments, including Human Snail Alexa Fluor ® 488-conjugated Antibody, Cat# IC3639G; Human/Mouse/Rat Vimentin APC-conjugated Antibody, Cat# IC2105A; Human Fibronectin PE-conjugated Antibody, Cat# IC1918P; Human/Mouse Wnt-5a Antibody (Cat# MAB645); Human IL-27 PE-conjugated Antibody, Cat# IC25261P; Human BAFF/BLyS/TNFSF13B PE-conjugated Antibody, Cat# IC1241P, Human IL-17RA/IL-17R Antibody; Cat# MAB177; Mouse F(ab)2 IgG (H+L) Fluorescein-conjugated Antibody, Cat# F0103B; Rabbit IgG Fluorescein-conjugated Antibody, Cat# F0112; and Goat IgG (H+L) Fluorescein-conjugated Antibody, Cat# F0109, were purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Expressing, Flow Cytometry, Co-Culture Assay, Fluorescence

Journal: Acta Pharmaceutica Sinica. B

Article Title: Spinal astrocyte-derived interleukin-17A promotes pain hypersensitivity in bone cancer mice

doi: 10.1016/j.apsb.2024.09.016

Figure Lengend Snippet: The distribution of IL-17RA in the spinal cord dorsal horn. (A) Double immunofluorescence shows that IL-17RA-IR are predominantly co-localized with NeuN, β Ⅲ-tubulin (neuronal marker), VGLUT2 (excitatory neuronal marker), and PAX2 (inhibitory neuronal marker) in the spinal dorsal horn. Scale bar: 25 μm. (B) The proportion of IL-17RA-IR in different types of neurons in the spinal dorsal horn ( n = 5–10 mice). (C) Double immunofluorescence shows that a few IL-17RA-IR are co-localized with GFAP, but not IBA-1, in the spinal dorsal horn. Scale bar: 25 μm. (D, E) Western blot analysis reveals a significant increase in IL-17RA protein on PTD 21 and 28. ∗ P < 0.05, ∗∗ P < 0.01; two-way ANOVA ( n = 4–7 mice). Data are presented as the mean ± SEM. Sample sizes for details are indicated in bars.

Article Snippet: We purchased the mouse IL-17A antibody (sc-374218) and control IgG (sc-2025) from Santa Cruz, the mouse IL-17RA antibody (MAB4481) and control IgG (MAB006) from R&D system, and the rabbit CX3CR1 antibody (TP501) and control IgG (PP501P) from Torrey Pines Biolabs.

Techniques: Immunofluorescence, Marker, Western Blot

Journal: Acta Pharmaceutica Sinica. B

Article Title: Spinal astrocyte-derived interleukin-17A promotes pain hypersensitivity in bone cancer mice

doi: 10.1016/j.apsb.2024.09.016

Figure Lengend Snippet: Blockade or knockdown of IL-17RA in spinal dorsal horn neurons alleviates or prevents bone cancer pain. (A) Schematic of the protocol in experiments B. (B) Optogenetic activation-induced decreases in PWL and PWT in the ipsilateral hindpaw to light stimulation was prevented by pre i.t. injection of IL-17RA antibody (10 μg). ∗ P < 0.05, ∗∗ P < 0.01 versus control IgG; two-way ANOVA ( n = 4). (C) Schematic of the protocol in experiments D. (D) i.t. injection of IL-17RA neutralizing antibody (IL-17RA Ab, 10 μg) reverses bone cancer-induced thermal hyperalgesia and mechanical allodynia. ∗ P < 0.05, ∗∗ P < 0.01 versus control IgG; two-way RM ANOVA ( n = 6 mice). (E) Schematic of the protocol in experiments I and J. (F) Immunofluorescence staining confirms knockdown efficiency of IL-17RA-shRNA in dorsal horn neurons, as evidenced by few IL-17RA-IR co-localized with Syn-IL-17RA-shRNA-EGFP in dorsal horn neurons. Scale bar: 25 μm. (G) RNAscope FISH showing reduced Il-17r mRNA level by IL-17RA-shRNA in the spinal dorsal horn. Scale bar: 25 μm. ∗ P < 0.05; two-tailed Student's t -test ( n = 10 and 5 mice). (H) Western blot analysis showing reduced IL-17RA protein by knockdown of IL-17RA in the spinal dorsal horn. ∗ P < 0.05; two-tailed Student's t -test ( n = 4 and 6 mice). (I, J) Selective knockdown of IL-17RA in dorsal horn neurons prevents thermal hyperalgesia (I) and attenuates mechanical allodynia (J). ∗ P < 0.05, ∗∗ P < 0.01 versus Ctrl-shRNA; two-way RM ANOVA ( n = 11 and 13 mice). Data are presented as the mean ± SEM. Sample sizes for details are indicated in bars.

Article Snippet: We purchased the mouse IL-17A antibody (sc-374218) and control IgG (sc-2025) from Santa Cruz, the mouse IL-17RA antibody (MAB4481) and control IgG (MAB006) from R&D system, and the rabbit CX3CR1 antibody (TP501) and control IgG (PP501P) from Torrey Pines Biolabs.

Techniques: Knockdown, Activation Assay, Injection, Control, Immunofluorescence, Staining, shRNA, RNAscope, Two Tailed Test, Western Blot

Journal: Acta Pharmaceutica Sinica. B

Article Title: Spinal astrocyte-derived interleukin-17A promotes pain hypersensitivity in bone cancer mice

doi: 10.1016/j.apsb.2024.09.016

Figure Lengend Snippet: IL-17RA mediates bone cancer pain through CaMKII α phosphorylation in the spinal dorsal horn. (A) Immunofluorescence triple labeling shows that both VGLUT2 + and PAX2 + neurons express pCaMKII α in the spinal dorsal horn from bone cancer mice. Scale bar: 25 μm. (B) Western blot analysis shows that bone cancer upregulates pCaMKII α expression, which can be blocked by knockdown of IL-17RA in the spinal dorsal horn on PTD 21. ∗ P < 0.05, ∗∗ P < 0.01; one-way ANOVA ( n = 4–6 mice). (C, D) i.t. injection of CaMKII α inhibitor KN93 (45 nmoL/5 μL) attenuated bone cancer-induced thermal hyperalgesia (C) and mechanical allodynia (D). ∗∗ P < 0.01 versus KN92, ## P < 0.01 versus vehicle; two-way RM ANOVA ( n = 17–12 mice). (E) Immunofluorescence staining shows AAV2/9-GFAP-IL-17A-EGFP co-localized with IL-17A-IR in the dorsal horn. Scale bar: 25 μm. (F, G) Overexpression of IL-17A (IL-17A OE) in spinal astrocytes induces thermal hyperalgesia (F) and mechanical allodynia (G), which can be rescued by i.t. injection of KN93. ∗ P < 0.05, ∗∗ P < 0.01; two-way ANOVA ( n = 6–8 mice). Data are presented as the mean ± SEM. Sample sizes for details are indicated in bars and brackets.

Article Snippet: We purchased the mouse IL-17A antibody (sc-374218) and control IgG (sc-2025) from Santa Cruz, the mouse IL-17RA antibody (MAB4481) and control IgG (MAB006) from R&D system, and the rabbit CX3CR1 antibody (TP501) and control IgG (PP501P) from Torrey Pines Biolabs.

Techniques: Immunofluorescence, Labeling, Western Blot, Expressing, Knockdown, Injection, Staining, Over Expression

Journal: Acta Pharmaceutica Sinica. B

Article Title: Spinal astrocyte-derived interleukin-17A promotes pain hypersensitivity in bone cancer mice

doi: 10.1016/j.apsb.2024.09.016

Figure Lengend Snippet: Knockdown of IL-17RA in spinal excitatory and inhibitory neurons, but not astrocytes, alleviates bone cancer pain. (A) Schematic of the protocol in experiments B−D. (B) Immunofluorescence staining confirms the knockdown efficiency of IL-17RA-shRNA in Vglut2 + excitatory neurons, as evidenced by few IL-17RA-IR co-localized with DIO-IL-17RA-shRNA-EGFP in a Vglut2 -Cre mouse. Scale bar: 25 μm. (C, D) Knockdown of IL-17RA in dorsal horn Vglut2 + excitatory neurons delays and attenuates bone cancer-induced thermal hyperalgesia (C) and mechanical allodynia (D). ∗ P < 0.05, ∗∗ P < 0.01 versus Ctrl-shRNA; two-way RM ANOVA ( n = 6 and 7 mice). (E) Schematic of the protocol in experiments G and H. (F) Immunofluorescence staining confirms knockdown efficiency of IL-17RA-shRNA in Vgat + inhibitory neurons, few IL-17RA-IR co-localized with DIO-IL-17RA-shRNA-EGFP in a Vgat -Cre mouse. Scale bar: 25 μm. (G, H) Knockdown of IL-17RA in dorsal horn Vgat + inhibitory neurons blocks and attenuates bone cancer-induced thermal hyperalgesia (G) and mechanical allodynia (H). ∗ P < 0.05, ∗∗ P < 0.01 versus Ctrl-shRNA; two-way RM ANOVA ( n = 6 and 7 mice). (I) Schematic of the protocol in experiments K and L. (J) Immunofluorescence staining confirms knockdown efficiency of IL-17RA-shRNA in astrocytes, few IL-17RA-IR co-localized with GfaABC1D-IL-17RA-shRNA-EGFP in the spinal dorsal horn. Scale bar: 25 μm. (K, L) Selective knockdown of IL-17RA in dorsal horn astrocytes does not affect the development of bone cancer pain. Two-way RM ANOVA ( n = 6 mice). Data are presented as the mean ± SEM. Sample sizes for details are indicated in brackets.

Article Snippet: We purchased the mouse IL-17A antibody (sc-374218) and control IgG (sc-2025) from Santa Cruz, the mouse IL-17RA antibody (MAB4481) and control IgG (MAB006) from R&D system, and the rabbit CX3CR1 antibody (TP501) and control IgG (PP501P) from Torrey Pines Biolabs.

Techniques: Knockdown, Immunofluorescence, Staining, shRNA

Journal: International immunopharmacology

Article Title: MLN4924 alleviates autoimmune myocarditis by promoting Act1 degradation and blocking Act1-mediated mRNA stability.

doi: 10.1016/j.intimp.2024.112716

Figure Lengend Snippet: Fig. 5. MLN4924 disrupts IL-17R/Act1/TRAF6 complex formation. (A) Immunoprecipitation of Act1 and western blot analysis of Act1, TRAF6, and IL-17R in H9C2 cells treated with 50 ng/ml rat recombinant IL-17A with or without 5 μM MLN4924 for 4 h. The bar graph represents the statistical analysis of protein expression. (B) Western blot analysis of nuclear Act1 protein concentration in H9C2 cells after treatment with IL-17A and/or MLN4924 for 4 h. The bar graph represents the statistical analysis of nuclear Act1 protein expression. The gray values of western blot bands were quantified using Image J software. The data are shown as means ± SD, n = 3. MLN: MLN4924.

Article Snippet: Protein A/G agarose (Cat#sc-2003) and antibodies against IL-17R (Cat#sc-376374, 1:200), TRAF6 (Cat#sc-8409, 1:200), Act1 (Cat#sc-398161, 1:200), and mouse IgG (Cat#sc-3877) were obtained from Santa Cruz Biotechnology.

Techniques: Immunoprecipitation, Western Blot, Recombinant, Expressing, Protein Concentration, Software