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Antibodies used in this study

Journal: Endocrinology

Article Title: Roles of Growth Hormone–Dependent JAK-STAT5 and Lyn Kinase Signaling in Determining Lifespan and Cancer Incidence

doi: 10.1210/endocr/bqae136

Figure Lengend Snippet: Antibodies used in this study

Article Snippet: P-p38 MAPK (Thr180/Tyr182) , 4511 , AB 2139682 , Cell Signaling Technology.

Techniques:

Altered hepatic transcripts involved in inflammatory stimuli in 12-week-old Ghr -mutant mice fed ad libitum. (A) Elevated transcripts of IL-10rα and IL-10rβ in male Ghr mutant and interleukin 1 receptor antagonist ( IL-1Ra ), cyclin-dependent kinase inhibitor 1A ( Cdkn1a ), in Ghr −/− males and females. No change was evident in IL-1β levels. (B) Decreased protein levels of inflammation modulators in livers of 12-week-old male Ghr mutants fed ad libitum. Decrease expression of acetylated tubulin (K40), phosphorylated p38 stress kinase, as well as inflammatory and immune response modulator nuclear factor (NF)κB p65 subunit and oxidative stress generator NADPH oxidase 4 (Nox4) in Ghr −/− and GhrBox1 −/− mice. Blots representative of n = 3 mice/genotype. Data represented as mean ± SEM (n = 4 mice/genotype). * P less than .05; ** P less than .01; *** P less than .001; **** P less than .0001 compared to wild-type (WT) of the same sex by analysis of variance.

Journal: Endocrinology

Article Title: Roles of Growth Hormone–Dependent JAK-STAT5 and Lyn Kinase Signaling in Determining Lifespan and Cancer Incidence

doi: 10.1210/endocr/bqae136

Figure Lengend Snippet: Altered hepatic transcripts involved in inflammatory stimuli in 12-week-old Ghr -mutant mice fed ad libitum. (A) Elevated transcripts of IL-10rα and IL-10rβ in male Ghr mutant and interleukin 1 receptor antagonist ( IL-1Ra ), cyclin-dependent kinase inhibitor 1A ( Cdkn1a ), in Ghr −/− males and females. No change was evident in IL-1β levels. (B) Decreased protein levels of inflammation modulators in livers of 12-week-old male Ghr mutants fed ad libitum. Decrease expression of acetylated tubulin (K40), phosphorylated p38 stress kinase, as well as inflammatory and immune response modulator nuclear factor (NF)κB p65 subunit and oxidative stress generator NADPH oxidase 4 (Nox4) in Ghr −/− and GhrBox1 −/− mice. Blots representative of n = 3 mice/genotype. Data represented as mean ± SEM (n = 4 mice/genotype). * P less than .05; ** P less than .01; *** P less than .001; **** P less than .0001 compared to wild-type (WT) of the same sex by analysis of variance.

Article Snippet: P-p38 MAPK (Thr180/Tyr182) , 4511 , AB 2139682 , Cell Signaling Technology.

Techniques: Mutagenesis, Expressing

PI3K/AKT/mTOR and MAPK/ERK1/2 signaling pathways may be involved in the therapeutic effect of HLJDD on PAAD. Western blot analysis of protein levels of p-mTOR, mTOR, p-AKT, AKT, p-PI3K, PI3K, p38 MAPK, p-p38 MAPK, ERK1/2, p-ERK1/2 and GAPDH in Capan-1 and Panc02 cells. All data were expressed as mean ± SD (ns p > .05; * p < .05; ** p < .01; *** p < .001; **** p < .0001).

Journal: Journal of Evidence-based Integrative Medicine

Article Title: “Huanglianjiedu Decoction” Against Pancreatic Adenocarcinoma Proliferation of by Downregulating the PI3K/AKT/mTOR and MAPK/ERK1/2 Signaling Pathways

doi: 10.1177/2515690X241291381

Figure Lengend Snippet: PI3K/AKT/mTOR and MAPK/ERK1/2 signaling pathways may be involved in the therapeutic effect of HLJDD on PAAD. Western blot analysis of protein levels of p-mTOR, mTOR, p-AKT, AKT, p-PI3K, PI3K, p38 MAPK, p-p38 MAPK, ERK1/2, p-ERK1/2 and GAPDH in Capan-1 and Panc02 cells. All data were expressed as mean ± SD (ns p > .05; * p < .05; ** p < .01; *** p < .001; **** p < .0001).

Article Snippet: The primary antibodies used included anti-Ki-67 antibody (Cat. #ab15580, Abcam, UK), anti-p-ERK1/2 antibody (Cat. #4370, Cell Signaling Technology, CST, USA), and anti-p-p38 MAPK antibody (Thr180/Tyr182) (Cat. #4511S, CST).

Techniques: Western Blot

Histological and immunohistochemical analysis of tumor tissues from C57BL/6 mice with subcutaneous Panc02 xenografts treated with HLJDD. Hematoxylin and Eosin (HE) staining illustrates the tumor architecture at 1× and 10× magnification across control (vehicle), 50 mg/kg, 100 mg/kg, and 200 mg/kg HLJDD-treated groups. Ki-67 immunohistochemistry staining, shown at 1× and 10× magnification, indicates cell proliferation, with reduced staining intensity in HLJDD-treated groups, suggesting decreased proliferation. Phosphorylated p38 MAPK (p-p38 MAPK) and phosphorylated ERK (p-ERK) staining at 1× and 10× magnification show decreased staining intensity with increasing HLJDD doses, indicating inhibition of the MAPK and ERK signaling pathways. Scale bars represent 2.5 mm (1×) and 200 μm (10×).

Journal: Journal of Evidence-based Integrative Medicine

Article Title: “Huanglianjiedu Decoction” Against Pancreatic Adenocarcinoma Proliferation of by Downregulating the PI3K/AKT/mTOR and MAPK/ERK1/2 Signaling Pathways

doi: 10.1177/2515690X241291381

Figure Lengend Snippet: Histological and immunohistochemical analysis of tumor tissues from C57BL/6 mice with subcutaneous Panc02 xenografts treated with HLJDD. Hematoxylin and Eosin (HE) staining illustrates the tumor architecture at 1× and 10× magnification across control (vehicle), 50 mg/kg, 100 mg/kg, and 200 mg/kg HLJDD-treated groups. Ki-67 immunohistochemistry staining, shown at 1× and 10× magnification, indicates cell proliferation, with reduced staining intensity in HLJDD-treated groups, suggesting decreased proliferation. Phosphorylated p38 MAPK (p-p38 MAPK) and phosphorylated ERK (p-ERK) staining at 1× and 10× magnification show decreased staining intensity with increasing HLJDD doses, indicating inhibition of the MAPK and ERK signaling pathways. Scale bars represent 2.5 mm (1×) and 200 μm (10×).

Article Snippet: The primary antibodies used included anti-Ki-67 antibody (Cat. #ab15580, Abcam, UK), anti-p-ERK1/2 antibody (Cat. #4370, Cell Signaling Technology, CST, USA), and anti-p-p38 MAPK antibody (Thr180/Tyr182) (Cat. #4511S, CST).

Techniques: Immunohistochemical staining, Staining, Control, Immunohistochemistry, Inhibition

Rescue experiment demonstrating the effects of HLJDD on the PI3K/AKT/mTOR and MAPK signaling pathways in Capan-1 and Panc02 cells. (A) Immunoblot analysis of PI3K, p-PI3K, AKT, p-AKT, mTOR, and p-mTOR in Capan-1 and Panc02 cells treated with HLJDD (200 μg/mL) in the presence or absence of the PI3K activator 740 Y-P (30 μM). HLJDD treatment reduces the levels of phosphorylated PI3K, AKT, and mTOR, indicating inhibition of the PI3K/AKT/mTOR pathway. The addition of 740 Y-P partially reverses this inhibition, restoring phosphorylation levels. (B) Immunoblot analysis of p38 MAPK, p-p38 MAPK, ERK1/2, and p-ERK1/2 in Capan-1 and Panc02 cells treated with HLJDD (200 μg/mL) in the presence or absence of the PKC activator PMA (500 nM). HLJDD treatment reduces the levels of phosphorylated p38 MAPK and ERK1/2, indicating inhibition of the MAPK pathway. The addition of PMA partially reverses this inhibition, restoring phosphorylation levels. GAPDH serves as a loading control in all experiments.

Journal: Journal of Evidence-based Integrative Medicine

Article Title: “Huanglianjiedu Decoction” Against Pancreatic Adenocarcinoma Proliferation of by Downregulating the PI3K/AKT/mTOR and MAPK/ERK1/2 Signaling Pathways

doi: 10.1177/2515690X241291381

Figure Lengend Snippet: Rescue experiment demonstrating the effects of HLJDD on the PI3K/AKT/mTOR and MAPK signaling pathways in Capan-1 and Panc02 cells. (A) Immunoblot analysis of PI3K, p-PI3K, AKT, p-AKT, mTOR, and p-mTOR in Capan-1 and Panc02 cells treated with HLJDD (200 μg/mL) in the presence or absence of the PI3K activator 740 Y-P (30 μM). HLJDD treatment reduces the levels of phosphorylated PI3K, AKT, and mTOR, indicating inhibition of the PI3K/AKT/mTOR pathway. The addition of 740 Y-P partially reverses this inhibition, restoring phosphorylation levels. (B) Immunoblot analysis of p38 MAPK, p-p38 MAPK, ERK1/2, and p-ERK1/2 in Capan-1 and Panc02 cells treated with HLJDD (200 μg/mL) in the presence or absence of the PKC activator PMA (500 nM). HLJDD treatment reduces the levels of phosphorylated p38 MAPK and ERK1/2, indicating inhibition of the MAPK pathway. The addition of PMA partially reverses this inhibition, restoring phosphorylation levels. GAPDH serves as a loading control in all experiments.

Article Snippet: The primary antibodies used included anti-Ki-67 antibody (Cat. #ab15580, Abcam, UK), anti-p-ERK1/2 antibody (Cat. #4370, Cell Signaling Technology, CST, USA), and anti-p-p38 MAPK antibody (Thr180/Tyr182) (Cat. #4511S, CST).

Techniques: Western Blot, Inhibition, Control