pdk1 primary antibody  (Proteintech)

Journal: CytoJournal

Article Title: Nobiletin alleviates brain injury in uremic mice and inhibits indoxyl sulfate-induced neurotoxicity in HT22 cells through the phosphatidylinositol 3-kinase/protein kinase B signaling pathway

doi: 10.25259/Cytojournal_233_2024

Figure Lengend Snippet: NOB alleviates DDP-induced brain damage in uremic mice. (a) Body weight of mice. (b) Kidney coefficient of mice. (c-f) Serum levels of BUN, SCr, Kim-1, and Nagl in mice. (g) Representative images of hematoxylin & eosin staining morphological analysis of kidney tissue, showing renal injury features including cellular debris, tubular necrosis, and inflammatory cells ×100. Scale bar = 100 μm (h-k) WB analysis and quantification of Bcl-2, Bax, and PARP protein expression in mouse brain tissue. n = 3; ✶ ✶ ✶ P < 0.001, ✶ ✶ P < 0.01 versus Control, ### P < 0.001, ## P < 0.01, # P < 0.05 versus DDP. The bar represents mean ± S.D. NOB: Nobiletin, BUN: Blood urea nitrogen, SCr: Serum creatinine, Bcl2: B-cell lymphoma-2, Bax: Bcl-2-associated X protein, PARP: Poly ADP-ribose polymerase, DDP: Cisplatin, WB: Western blot, S.D.: Standard deviation.

Article Snippet: The membrane was then blocked using 5% nonfat milk at room temperature for 1 h and subsequently incubated overnight at 4°C with primary antibodies targeting α-tubulin (2144), Bcl2 (15071), poly ADP-ribose polymerase (PARP) (9532), PI3K (4292), pyruvate dehydrogenase kinase 1 (PDK1) (3062), Akt (4691), p-PI3K (17366), p-PDK1 (3438), p-Akt (4060), and Bax (5023) (1:1000 each, Cell Signaling Technology, Danvers, MA, USA).

Techniques: Staining, Expressing, Control, Western Blot, Standard Deviation

Journal: CytoJournal

Article Title: Nobiletin alleviates brain injury in uremic mice and inhibits indoxyl sulfate-induced neurotoxicity in HT22 cells through the phosphatidylinositol 3-kinase/protein kinase B signaling pathway

doi: 10.25259/Cytojournal_233_2024

Figure Lengend Snippet: NOB exhibits anti-apoptotic effects on DDP-induced HT22 cells. (a and b) CCK-8 assay evaluating the impact of DDP and NOB on HT22 cell viability. ✶ ✶ ✶ P < 0.0001, ✶ ✶ P < 0.001, ✶ P < 0.01 versus 0 (c) CCK-8 assay assessing the effect of various concentrations of NOB on DDP-induced HT22 cell viability. (d-g) WB analysis and quantification of Bcl-2, Bax, and PARP protein expression in HT22 cells across different groups. (h-i) DCFH-DA probe measurement of intracellular ROS levels in different groups. ×100 Scale bar = 100 μm. n = 3; ✶ ✶ ✶ P < 0.001, ✶ ✶ P < 0.01, ✶ P < 0.05 versus DDP; ### P < 0.001 versus Control. The bar represents mean ± S.D. NOB: Nobiletin, DDP: Cisplatin, CCK-8: Cell counting kit-8, Bcl2: B-cell lymphoma-2, Bax: Bcl-2-associated X protein, PARP: Poly ADP-ribose polymerase, WB: Western blot, DCFH-DA: Dichlorodihydrofluorescein diacetate, ROS: Reactive oxygen species.

Article Snippet: The membrane was then blocked using 5% nonfat milk at room temperature for 1 h and subsequently incubated overnight at 4°C with primary antibodies targeting α-tubulin (2144), Bcl2 (15071), poly ADP-ribose polymerase (PARP) (9532), PI3K (4292), pyruvate dehydrogenase kinase 1 (PDK1) (3062), Akt (4691), p-PI3K (17366), p-PDK1 (3438), p-Akt (4060), and Bax (5023) (1:1000 each, Cell Signaling Technology, Danvers, MA, USA).

Techniques: CCK-8 Assay, Expressing, Control, Cell Counting, Western Blot

Journal: CytoJournal

Article Title: Nobiletin alleviates brain injury in uremic mice and inhibits indoxyl sulfate-induced neurotoxicity in HT22 cells through the phosphatidylinositol 3-kinase/protein kinase B signaling pathway

doi: 10.25259/Cytojournal_233_2024

Figure Lengend Snippet: NOB inhibits apoptosis through the PI3K/Akt signaling pathway. (a-h) WB detection and quantitative analysis of the protein expression of p-PI3K, p-PDK1, and p-Akt in different groups of HT22 cells. (i-l) WB detection and quantitative analysis of the protein expression of Bcl2, Bax, and PARP in different groups of HT22 cells. (m and n) DCFH-DA probe measurement of intracellular ROS levels in different groups. ×100 Scale bar = 100 μm (o) CCK-8 assay assessing the effect of various LY concentrations on HT22 cell viability. n = 3; ✶ ✶ ✶ P < 0.001, ✶ ✶ P < 0.01, ✶ P < 0.05 versus DDP; ## P < 0.01, # P < 0.05 versus DDP + NOB. The bar represents mean ± S.D. NOB: Nobiletin, DDP: Cisplatin, PI3K: Phosphatidylinositol 3-kinase, PDK1: Pyruvate dehydrogenase kinase 1, Akt: Protein kinase B, Bcl2: B-cell lymphoma-2, Bax: Bcl-2-associated X protein, PARP: Poly ADP-ribose polymerase, WB: Western blot, DCFH-DA: Dichlorodihydrofluorescein diacetate, ROS: Reactive oxygen species, LY: LY294002, CCK-8: Cell counting kit-8.

Article Snippet: The membrane was then blocked using 5% nonfat milk at room temperature for 1 h and subsequently incubated overnight at 4°C with primary antibodies targeting α-tubulin (2144), Bcl2 (15071), poly ADP-ribose polymerase (PARP) (9532), PI3K (4292), pyruvate dehydrogenase kinase 1 (PDK1) (3062), Akt (4691), p-PI3K (17366), p-PDK1 (3438), p-Akt (4060), and Bax (5023) (1:1000 each, Cell Signaling Technology, Danvers, MA, USA).

Techniques: Expressing, CCK-8 Assay, Western Blot, Cell Counting