rabbit anti mouse inos antibody  (Proteintech)

Journal: Journal of Advanced Research

Article Title: A novel TGR5 agonist Sauchinone ameliorates IMQ induced murine psoriasis by regulating macrophage polarization

doi: 10.1016/j.jare.2025.04.034

Figure Lengend Snippet: Sauchinone inhibits M1 macrophage polarization and promotes M2 macrophage polarization in BMDM. (A) The cytotoxicity of Sauchinone on BMDM was detected by CCK-8 kit. (B-G) BMDM were stimulated with LPS plus IFN-γ to induce M1 polarization in the presence or absence of different doses of Sauchinone. The transcription levels of Tnf-α , Il-6 , Il −12 , Inos , Il-1β , Cox-2 and Nlrp3 (B) were detected by RT-qPCR. TNF-α, IL-6, IL-12 and NO levels in culture supernatants (C) were measured by ELISA or Griess assay. The protein levels of p-STAT-1, STAT-1, and INOS in cell lysates (D) were detected by western blot. The percentages of INOS + , MHCⅡ + and CD86 + BMDM (E) were measured by FACS. (F-H) BMDM were stimulated with IL-4 plus IL-13 to induce M2 polarization in the presence or absence of different doses of Sauchinone. The transcription levels of Arg-1, Fizz1 , Ym-1 , Cd206 , Cd301 , and Dectin-1 (F) were detected by RT-qPCR. The protein levels of p-STAT-6 and STAT-6 in cell lysates (G) were detected by western blot. The percentages of CD301 + BMDM (H) were measured by FACS. Data were shown as Mean ± SEM from triplicate measurements. *P < 0.05, **P < 0.01 compared as indicated.

Article Snippet: Anti-mouse STAT-1 (9712L), anti-mouse p-STAT-1 (7649L), anti-mouse Inducible nitric oxide synthase (INOS) (13120S), Anti-mouse STAT-6 (9362S), anti-mouse p-STAT-6 (56554S), anti-mouse PKA (5842S), anti-mouse p-CREB (9197S), anti-mouse CREB (9198L),anti-mouse p-IκB (9246L), anti-mouse IκB (4812S), anti-mouse p-p65 (3033L), anti-mouse p65 (4764S), anti-mouse ASC (67824S) and anti-mouse NLRP3 (15101S) were obtained from Cell Signaling Technology (Danvers, MA, USA).

Techniques: CCK-8 Assay, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Griess Assay, Western Blot

Journal: Journal of Advanced Research

Article Title: A novel TGR5 agonist Sauchinone ameliorates IMQ induced murine psoriasis by regulating macrophage polarization

doi: 10.1016/j.jare.2025.04.034

Figure Lengend Snippet: Sauchinone inhibits macrophage M1 polarization in TGR5 dependent manner. (A-B) BMDM from WT mice were stimulated with LPS plus IFN-γ to induce M1 polarization in the presence or absence of Sauchinone (30 μM) or SBI-115 (5 μM). The transcription levels of Tnf-α , Il-6 , Inos and Il-1β (A) were detected by RT-qPCR. The protein levels of p-STAT-1, STAT-1 and INOS in cell lysates (B) were detected by western blot. (C-E) BMDM from WT mice were stimulated with LPS plus IFN-γ to induce M1 polarization in the presence or absence of Sauchinone (30 μM) or TGR5 si-RNA. The transcription levels of Tnf-α , Il-6 , Inos and Il-1β (C) were detected by RT-qPCR. The protein levels of p-STAT-1, STAT-1 and INOS in cell lysates (D) were detected by western blot. IL-6 and NO levels in culture supernatants (E) were measured by ELISA and Griess assay. (F-H) BMDM from Tgr5 -/- mice were stimulated with LPS plus IFN-γ to induce M1 polarization in the presence or absence of Sauchinone (30 μM). IL-6 and NO levels in culture supernatants (F) were measured by ELISA or Griess assay. The transcription levels of Tnf-α , Il-6 , Inos and Il-1β (G) were detected by RT-qPCR. The protein levels of p-STAT-1, STAT-1 and INOS in cell lysates (H) were detected by western blot. Data were shown as Mean ± SEM from triplicate measurements. *P < 0.05, **P < 0.01 compared as indicated.

Article Snippet: Anti-mouse STAT-1 (9712L), anti-mouse p-STAT-1 (7649L), anti-mouse Inducible nitric oxide synthase (INOS) (13120S), Anti-mouse STAT-6 (9362S), anti-mouse p-STAT-6 (56554S), anti-mouse PKA (5842S), anti-mouse p-CREB (9197S), anti-mouse CREB (9198L),anti-mouse p-IκB (9246L), anti-mouse IκB (4812S), anti-mouse p-p65 (3033L), anti-mouse p65 (4764S), anti-mouse ASC (67824S) and anti-mouse NLRP3 (15101S) were obtained from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay, Griess Assay

Journal: Journal of Advanced Research

Article Title: A novel TGR5 agonist Sauchinone ameliorates IMQ induced murine psoriasis by regulating macrophage polarization

doi: 10.1016/j.jare.2025.04.034

Figure Lengend Snippet: Sauchinone inhibits macrophage M1 polarization through TGR5/PKA pathway. (A-C) BMDM were stimulated with LPS plus IFN-γ to induce M1 polarization in the presence or absence of different doses of Sauchinone (30 μM) or PKA inhibitor H89 (10 μM). IL-6 and NO levels in culture supernatants (A) were measured by ELISA or Griess assay. The transcription levels of Tnf-α , Il-6 , Inos and Il-1β (B) were detected by RT-qPCR. The protein levels of p-STAT-1, STAT-1 and INOS in cell lysates (C) were detected by western blot. Data were shown as Mean ± SEM from triplicate measurements. *P < 0.05, **P < 0.01 compared as indicated.

Article Snippet: Anti-mouse STAT-1 (9712L), anti-mouse p-STAT-1 (7649L), anti-mouse Inducible nitric oxide synthase (INOS) (13120S), Anti-mouse STAT-6 (9362S), anti-mouse p-STAT-6 (56554S), anti-mouse PKA (5842S), anti-mouse p-CREB (9197S), anti-mouse CREB (9198L),anti-mouse p-IκB (9246L), anti-mouse IκB (4812S), anti-mouse p-p65 (3033L), anti-mouse p65 (4764S), anti-mouse ASC (67824S) and anti-mouse NLRP3 (15101S) were obtained from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Griess Assay, Quantitative RT-PCR, Western Blot

Journal: Journal of Advanced Research

Article Title: A novel TGR5 agonist Sauchinone ameliorates IMQ induced murine psoriasis by regulating macrophage polarization

doi: 10.1016/j.jare.2025.04.034

Figure Lengend Snippet: Sauchinone decreases the markers of M1 macrophages and increases the markers of M2 macrophages in skin infiltrated macrophages. (A) The percentages of M1-like (CD86 + ) macrophage and M2-like macrophage (CD206 + or CD301 + ) in skin infiltrated macrophages (CD11b + , F4/80 + ) were detected by FACS. (b-d) The expression of M1-like markers CD86 (B) and INOS (C) and M2-like marker CD206 (D) on skin infiltrated macrophages (stained with F4/80) were detected by immunofluorescence. Scale bar: 200 µm.

Article Snippet: Anti-mouse STAT-1 (9712L), anti-mouse p-STAT-1 (7649L), anti-mouse Inducible nitric oxide synthase (INOS) (13120S), Anti-mouse STAT-6 (9362S), anti-mouse p-STAT-6 (56554S), anti-mouse PKA (5842S), anti-mouse p-CREB (9197S), anti-mouse CREB (9198L),anti-mouse p-IκB (9246L), anti-mouse IκB (4812S), anti-mouse p-p65 (3033L), anti-mouse p65 (4764S), anti-mouse ASC (67824S) and anti-mouse NLRP3 (15101S) were obtained from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Expressing, Marker, Staining, Immunofluorescence

Journal: Scientific Reports

Article Title: Barium titanate piezoelectric nanoparticles induce M1 polarization in mouse macrophages via ultrasound in vitro

doi: 10.1038/s41598-025-23364-6

Figure Lengend Snippet: ( A ) Fluorescence images of RAW264.7 cells under different conditions demonstrating bioelectrically-driven differentiation of M0 monocytes into M1 phenotype macrophages. Phase (Column 1), DAPI staining intensity (Column 2), and iNOS staining intensity (Column 3) are shown. Row 1 shows biochemical treatment with IFN-γ + LPS that drives M0 to M1. Row 2 shows positive iNOS intensity effect with pzNP and uS (+ NP, + uS). Rows 3, 4 and 5 represent controls with less/negligible iNOS staining intensity: with pzNPs and without uS (+ NP, –uS, Row 3), without pzNPs and with uS (–NP, + uS, Row 4), and without pzNPs or uS (-NP, -uS, Row 5). Note that the bioelectric combination of pzNP uptake and uS inducing iNOS expression in Row 2 is similar to biochemical treatment with IFN-γ + LPS in Row 1, both indicative of M0→M1 differentiation. ( B ) Average mean iNOS fluorescence intensity of cells within regions of interest from sets of n = 3 fluorescent images quantified using FIJI/ImageJ, for each NP/uS condition shown, plus that for IFN-γ + LPS. A p value > 0.1 (indicated as NS , for Not Significant) was determined via a t -test comparing the IFN-γ + LPS and (+ NP, + uS) data, while p < 0.01 (indicated as **) when comparing those with the three control data sets, as determined via ANOVA analysis. See text and Statistical Analysis for details.

Article Snippet: Then, a primary rabbit polyclonal antibody against mouse iNOS (Cell Signaling Technology, Danvers MA, no. 68186) or mouse arginase (Cell Signaling Technology, no. 93668), a marker for M1, was incubated overnight in PBS (1:500 dilution) at 4 °C.

Techniques: Fluorescence, Staining, Expressing, Control

Journal: Scientific Reports

Article Title: Barium titanate piezoelectric nanoparticles induce M1 polarization in mouse macrophages via ultrasound in vitro

doi: 10.1038/s41598-025-23364-6

Figure Lengend Snippet: Composite fluorescence images of a mixture of RAW264.7 macrophage cells exposed and not exposed to piezoelectric nanoparticle (pzNP) uptake, and then subjected to ultrasound (uS) stimulation . The channels displayed include phase contrast (panel A ), DAPI (blue, panel B ) for nuclear staining, iNOS expression (red, panel C ), and SA-FITC coating of pzNPs (green, panel D ), in cells subjected to pzNP uptake with uS stimulation. ( E ) A mixture of nanoparticle and non-nanoparticle loaded cells are detected. The results indicate that discrete subpopulations of cells exposed to pzNPs and uS can be identified. Cells that have taken up pzNPs but do not exhibit iNOS expression (green circle) can be differentiated from subpopulations that have taken up pzNPs and exhibit iNOS expression (white circle). There are subpopulations of cells that were not exposed to pzNPs and when subjected to ultrasound stimulation do not express iNOS expression (yellow circle).

Article Snippet: Then, a primary rabbit polyclonal antibody against mouse iNOS (Cell Signaling Technology, Danvers MA, no. 68186) or mouse arginase (Cell Signaling Technology, no. 93668), a marker for M1, was incubated overnight in PBS (1:500 dilution) at 4 °C.

Techniques: Fluorescence, Staining, Expressing