phosphorylated p pparγ  (Bioss)

Journal: International Journal of Oncology

Article Title: Cinobufagin inhibits M2-like tumor-associated macrophage polarization to attenuate the invasion and migration of lung cancer cells

doi: 10.3892/ijo.2024.5690

Figure Lengend Snippet: Effects of CB on the expression of macrophage polarization-related proteins. (A) Gel blots showing protein expression of Arg-1, iNOS, p-PPARγ and PPARγ in cells with different treatment groups. (B-D) Quantification of (B) Arg-1, (C) iNOS and (D) p-PPARγ based on the western blot assay (n=3). ** P<0.01 compared with M0 group; # P<0.05, ## P<0.01 and $$ P<0.01 compared with M2 group; ▲ P<0.05 and ▲▲ P<0.01 compared with M2 + RSG group. CB, cinobufagin; Arg-1, arginase 1; iNOS, inducible nitric oxide synthase; p-, phosphorylated; PPARγ, peroxisome proliferator-activated receptor γ; RSG, rosiglitazone; ns from M2 group, not statistically significant compared with M0 group; ns from M2 + RSG group, not statistically significant compared with M2 group.

Article Snippet: After that, primary antibodies against Arg-1 (1:1,000; cat. no. bs-23837R; BIOSS), inducible nitric oxide synthase (iNOS; 1:1,000; cat. no. bs-0162R; BIOSS); phosphorylated (p)-PPARγ (1:1,000; cat. no. bs-3737R; BIOSS), vimentin (1:1,000; cat. no. bsm-33170M; BIOSS), E-cadherin (1:1,000; cat. no. bs-1519R; BIOSS), N-cadherin (1:1,000; cat. no. bs-20623R; BIOSS) and GAPDH (1:3,000; cat. no. 60004-1-Ig; Proteintech Group, Inc.) were incubated with the membrane overnight at 4°C, followed by washing with Tris-buffered saline containing 0.05% Tween 20 and incubation with horseradish peroxidase-conjugated secondary antibodies goat anti-mouse IgG (1:10,000; cat. no. SA00001-1) and goat anti-rabbit IgG (1:10,000; SA00001-2; both from Proteintech Group, Inc.) at room temperature for 1 h. The western blot bands were visualized using an ECL kit (cat. no. PE0010; Beijing Solarbio Science & Technology Co., Ltd.).

Techniques: Expressing, Western Blot

Journal: International Journal of Molecular Medicine

Article Title: Adropin attenuates pancreatitis-associated lung injury through PPARγ phosphorylation-related macrophage polarization

doi: 10.3892/ijmm.2023.5298

Figure Lengend Snippet: Excessive M1 macrophage polarization is observed in the Adro-KO + L-Arg group. (A) Western blot analysis of lung tissue form the Adro-KO + L-Arg group. (B) Co-expression of CD68 and iNOS in lung tissue in L-Arg group determined using immunofluorescence. (C) Co-expression of CD68 and CD206 of lung tissue in the L-Arg group determined using immunofluorescence. (D) iNOS mRNA level in lung tissue from the Adro-KO + L-Arg group (n≥8). (E) CD68 mRNA level in lung tissue from the Adro-KO + L-Arg group (n≥7). (F) CD163 mRNA level in lung tissue from the Adro-KO + L-Arg group (n≥8). (G) Arg-1 mRNA level in lung tissue from the Adro-KO + L-Arg group (n≥8). (H) Quantitative analysis of immunofluorescence (CD68 + iNOS) staining (n≥5). (I) Quantitative analysis of immunofluorescence (CD68; CD206) staining in macrophages (n≥9). (J) Quantitative analysis of the western blots (PPARγ; n≥6); (K) Quantitative analysis of the western blots (PPARγ Ser273; n≥6). (L) Quantitative analysis of the western blots (PPARγ Ser112; n≥6). * P<0.05. L-Arg, L-arginine; NS, normal saline; Adro-KO, adropin knockout; iNOS, inducible nitric oxide synthase.

Article Snippet: The primary antibodies were used at 4°C overnight, and the antibody included Adropin (1:1,000, cat. no. PA5-72781, Thermo Fisher Scientific, Inc.); GADPH (1:10,000; cat. no. AC001, ABclonal); peroxisome proliferator-activated receptor γ (PPARγ; 1:1,000, cat. no. bsm-52220R, BIOSS); p-PPARγ Ser112 (1:1,000, cat. no. bs-3737R, BIOSS); p-PPARγ Ser273 (1:1,000, cat. no. bs-2875R, BIOSS), caspase-3 (1:1,000, cat. no. YC0006, ImmunoWay Biotechnology Company) and PARP1 (1:1,000, cat. no. A0942, ABclonal).

Techniques: Western Blot, Expressing, Immunofluorescence, Staining, Saline, Knock-Out

Journal: International Journal of Molecular Medicine

Article Title: Adropin attenuates pancreatitis-associated lung injury through PPARγ phosphorylation-related macrophage polarization

doi: 10.3892/ijmm.2023.5298

Figure Lengend Snippet: Adropin exogenous supplement induces M2 macrophage polarization. (A) Western blot analysis of lung tissue from the Adro-KO + L-Arg + Adro (34−76) group. (B) Quantitative analysis of the western blots (PPARγ, PPARγ Ser112, PPARγ Ser273) (n≥5). (C) Co-expression of CD68 and iNOS in lung tissue from the Adro-KO + L-Arg+Adro (34−76) group. (D) Co-expression of CD68 and iNOS in lung tissue from the Adro-KO + L-Arg + Adro (34−76) group. (E) Ratio of CD68/DAPI of lung in Adro-KO+ L-Arg+Adro (34−76) group(n≥3); (F) The ratio of iNOS/DAPI in lung tissue from the Adro-KO + L-Arg + Adro (34−76) group (n≥3). (G) Ratio of iNOS/CD68 in lung tissue from the Adro-KO + L-Arg + Adro (34−76) group (n≥3). (H) Ratio of CD68/DAPI in lung tissue from the Adro-KO + L-Arg + Adro (34−76) group (n≥3). (I) Ratio of CD206/DAPI in lung tissue from the Adro-KO + L-Arg + Adro (34−76) group (n≥3). (J) Ratio of CD206/CD68 in lung tissue from the Adro-KO + L-Arg + Adro (34−76) group (n≥3). (K) CD163 mRNA level in lung tissue from the Adro-KO + L-Arg + Adro (34−76) group (n≥6). (L) Arg-1 mRNA level in lung tissue from the Adro-KO + L-Arg + Adro (34−76) group (n≥6). (M) iNOS mRNA level in lung tissue from the Adro-KO + L-Arg + Adro (34−76) group (n≥6). (N) CD86 mRNA level level in lung tissue from the Adro-KO + L-Arg + Adro (34−76) group (n≥6). * P<0.05 and *** P<0.001. L-Arg, L-arginine; Adro-KO, adropin knockout; NS, normal saline; iNOS, inducible nitric oxide synthase; Arg-1, arginase 1.

Article Snippet: The primary antibodies were used at 4°C overnight, and the antibody included Adropin (1:1,000, cat. no. PA5-72781, Thermo Fisher Scientific, Inc.); GADPH (1:10,000; cat. no. AC001, ABclonal); peroxisome proliferator-activated receptor γ (PPARγ; 1:1,000, cat. no. bsm-52220R, BIOSS); p-PPARγ Ser112 (1:1,000, cat. no. bs-3737R, BIOSS); p-PPARγ Ser273 (1:1,000, cat. no. bs-2875R, BIOSS), caspase-3 (1:1,000, cat. no. YC0006, ImmunoWay Biotechnology Company) and PARP1 (1:1,000, cat. no. A0942, ABclonal).

Techniques: Western Blot, Expressing, Knock-Out, Saline

Journal: Life Science Alliance

Article Title: Hepatic DKK1-driven steatosis is CD36 dependent

doi: 10.26508/lsa.202201665

Figure Lengend Snippet: Antibodies used for Western blot and IHC.

Article Snippet: (p)-PPARγ antibody bs-3737R , Bioss.

Techniques: Western Blot