anti-phospho-unc-51-like autophagy-activating kinase-1 (ulk1) (ser757, #14202)  (Cell Signaling Technology Inc)

Journal: World Journal of Gastroenterology

Article Title: Chaperonin-containing tailless complex polypeptide 1 subunit 6A negatively regulates autophagy and protects colorectal cancer cells from cisplatin-induced cytotoxicity

doi: 10.3748/wjg.v31.i18.105729

Figure Lengend Snippet: Chaperonin-containing tailless complex polypeptide 1 subunit 6A affects the autophagy pathway in colorectal cancer cells. A and B: Cell lysates were extracted from knockout (KO)-Control (Ctrl) and KO-chaperonin-containing tailless complex polypeptide 1 subunit 6A (CCT6A) HT29 cells, as well as overexpression (OE)-Ctrl and OE-CCT6A SW480 cells, and then subjected to immunoblotting using the indicated antibodies; C: Following treatment with chloroquine diphosphate salt (CQ, 20 μM, hereafter unless otherwise indicated) for 2 hours, immunoblotting was performed with the total proteins extracted from the indicated cells. DDP: Cisplatin; LC3: Light chain 3; mTOR: Mammalian target of rapamycin; Ulk1: Unc-51-like autophagy activating kinase 1.

Article Snippet: Primary antibodies against CCT6A (19793-1-AP), tubulin (66240-1-Ig), phosphorylated Unc-51-like autophagy activating kinase 1 (phospho-Ulk1) (Ser556, 80218-1-RR), Beclin-1 (11306-1-AP), and p62 (66184-1-Ig) were purchased from Proteintech Group (Wuhan, China).

Techniques: Knock-Out, Control, Over Expression, Western Blot

Journal: Advances in clinical and experimental medicine : official organ Wroclaw Medical University

Article Title: Icariin ameliorates osteoporosis by activating autophagy in ovariectomized rats.

doi: 10.17219/acem/174078

Figure Lengend Snippet: Fig. 6. Icariin mediated autophagy pathways bidirectionally. A. The expression of p-serine/ threonine protein kinase (Akt)/ Akt total, p-mammalian target of rapamycin (mTOR)/mTOR total, p-AMP-activated protein kinase (AMPK)/AMPK total, p-unc- 51-like autophagy activating kinase 1 (ULK1) (Ser555)/ULK1 total, and p-ULK (Ser757)/ULK1 total using western blotting; B. Quantification of p-Akt/ Akt total (H = 6.25, p = 0.04); C. Quantification of p-mTOR/ mTOR total (H = 6.25, p = 0.04); D. Quantification of p-AMPK/ AMPK total (H = 6.30, p = 0.04); E. Quantification of p-ULK1 (Ser555)/ULK1 total (H = 6.25, p = 0.04); F. Quantification of p-ULK1 (Ser 757)/ULK1 total (H = 6.25, p = 0.04). The Kruskal– Wallis H test and Dunn’s post hoc test were performed in the B, C, D, E, and F plots. Box bodies represent M (p25, p75), and upper bars and lower bars represent maximum and minimum, respectively (n = 3). The p-AKT/ AKT, p-mTOR/mTOR and p-ULK1 (Ser757)/ULK1 were increased, while p-AMPK/AMPK and p-ULK1 (Ser555)/ULK1 were decreased in the model group. Icariin reduced p-AKT/AKT, p-mTOR/ mTOR and p-ULK1 (Ser757)/ULK1, and increased p-AMPK/AMPK and p-ULK1 (Ser555)/ULK1Fig. 6. Icariin mediated autophagy pathways bidirectionally. A. The expression of p-serine/threonine protein kinase (Akt)/Akt total, p-mammalian target of rapamycin (mTOR)/mTOR total, p-AMP-activated protein kinase (AMPK)/AMPK total, p-unc-51-like autophagy activating kinase 1 (ULK1) (Ser555)/ULK1 total, and p-ULK (Ser757)/ULK1 total using western blotting; B. Quantification of p-Akt/Akt total (H = 6.25, p = 0.04); C. Quantification of p-mTOR/mTOR total (H = 6.25, p = 0.04); D. Quantification of p-AMPK/AMPK total (H = 6.30, p = 0.04); E. Quantification of p-ULK1 (Ser555)/ULK1 total (H = 6.25, p = 0.04); F. Quantification of p-ULK1 (Ser 757)/ULK1 total (H = 6.25, p = 0.04). The Kruskal–Wallis H test and Dunn’s post hoc test were performed in the B, C, D, E, and F plots. Box bodies represent M (p25, p75), and upper bars and lower bars represent maximum and minimum, respectively (n = 3). The p-AKT/AKT, p-mTOR/mTOR and p-ULK1 (Ser757)/ULK1 were increased, while p-AMPK/AMPK and p-ULK1 (Ser555)/ULK1 were decreased in the model group. Icariin reduced p-AKT/AKT, p-mTOR/mTOR and p-ULK1 (Ser757)/ULK1, and increased p-AMPK/AMPK and p-ULK1 (Ser555)/ULK1

Article Snippet: Autophagy-related protein 7 (Atg7) (8558), Akt (9272), p-Akt (4058), Beclin1 (3495), mammalian target of rapamycin (mTOR) (2972), p-unc-51-like autophagy activating kinase 1 (ULK1) (Ser555) (5869), p-mTOR (2971), ULK1 (8054), p-ULK1 (Ser757) (14202), and AMPactivated protein kinase (AMPK) (5832) were purchased from Cell Signaling Technology (Danvers, USA).

Techniques: Expressing, Western Blot