rabbit anti p mtor cell signaling technology 2971s wb  (Cell Signaling Technology Inc)

Journal: Cellular and molecular gastroenterology and hepatology

Article Title: The Protective Role of DDIT4 in Helicobacter pylori-induced Gastric Metaplasia Through Metabolic Regulation of Ferroptosis.

doi: 10.1016/j.jcmgh.2024.101448

Figure Lengend Snippet: Figure 2. Verification of DDIT4 expression and H. pylori colonization in the gastric tissues of Ddit4D/D and Ddit4-/- mice. (A) The qRT-PCR analysis indicating the mRNA levels of DDIT4 in the stomach tissues of Ddit4þ/þ and Ddit4-/- mice. (B) The ratio of p-mTOR to total mTOR protein levels in infected WT and Ddit4-knockout mice. (C) Silver staining showing H. pylori colonization in the gastric tissues of mice (Scale bar, 10 mm). (D) Colony-forming units (CFUs) per gram stomach in animals were evaluated for H. pylori colonization by culture at 4 days after sacrifice. (E) IHC scores of ATP4A and ATP4B in infected gastric tissues of Ddit4þ/þ and Ddit4-/- mice. (F) Immunofluorescence staining for MUC2 (left panel) or FABP1 (right panel) on infected gastric tissues from Ddit4þ/þ and Ddit4-/- mice (Scale bar, 20 mm). ns, no significance. *P < .05; **P < .01; and ***P < .001 were considered significant.

Article Snippet: After blocking the sections in 1% BSA for 30 3 February 2025 11:24 pm ce JO 48 Table 1.The Primary Antibodies Used in This Study Antibody Source Catalogue number Application Working dilution DDIT4 Abcam ab106356 WB, IHC, IF WB 1: 1000 IHC 1: 600 IF 1: 100 mTOR Cell Signaling Technology #2983 WB 1: 1000 p-mTOR Cell Signaling Technology #5566 WB 1: 1000 ALOX15 Abcam ab244205 WB, IHC WB 1: 1000 IHC 1: 500 HMOX1 ZenBio R24541 WB, IHC WB 1: 1000 IHC 1: 400 GPX4 Proteintech #67763-1-1g WB, IHC WB 1: 1000 IHC 1: 1000 ATP4A Abcam ab231729 WB, IF, IHC WB 1: 1000 IF 1: 100 IHC 1: 200 ATP4B ZenBio #120298 WB, IHC WB 1:1000 IHC 1: 200 GIF Santacruz sc-514523 IF 1:50 GSII, Alexa FluorTM 647 Invitrogen L32451 IF 1:800 CD44V9 Novus Biologicals NBP2-53204 IF 1:100 MUC2 Cell Signaling Technology #88686 IF 1: 100 FABP1 Cell Signaling Technology #13368 IF 1:100 gH2AX Cell Signaling Technology 9718 IF 1: 100 TFF2 Thermo Fisher PA5-80111 WB 1: 1000 GAPDH ZenBio R24404 WB 1: 1000 b-actin ZenBio R51031 WB 1: 2000 IF, immunofluorescence; IHC, immunohistochemistry; WB, Western blot.

Techniques: Expressing, Quantitative RT-PCR, Infection, Knock-Out, Silver Staining, Staining

Journal: Cellular and molecular gastroenterology and hepatology

Article Title: The Protective Role of DDIT4 in Helicobacter pylori-induced Gastric Metaplasia Through Metabolic Regulation of Ferroptosis.

doi: 10.1016/j.jcmgh.2024.101448

Figure Lengend Snippet: Figure 14. Clinical relevance of DDIT4 and ferroptosis in human gastric intestinal metaplasia tissues. (A) Western blot data for the expression levels of DDIT4, ALOX15, HMOX1 and GPX4 in human GIM and non-atrophic gastritis (GS) tissues. (B–C) IHC staining showing DDIT4 and GPX4 expression in human GS and GIM tissues. Representative images (Scale bar, 10 mm) shown in (B). Quantification of staining shown in (C). (D–E) C11-BODIPY and DCFH-DA staining measured with a fluo- rescence microscope showing lipid peroxidation (D) and ROS levels (E) in human GS and GIM tissues. (F) Diagram of the mechanism by which loss of DDIT4 promoted H. pylori-induced gastric metaplasia lesions through metabolic regulation of ferroptosis. Chronic H. pylori infection causes the loss of DDIT4 and mTOR signaling pathway activation, which in turn inhibits GSH synthesis and increases ROS levels and lipid peroxides, inducing ferroptosis in the gastric microenvironment. As a result, gastric metaplasia lesions develop following parietal cell loss. *P < .05; **P < .01; and ***P < .001 were considered significant.

Article Snippet: After blocking the sections in 1% BSA for 30 3 February 2025 11:24 pm ce JO 48 Table 1.The Primary Antibodies Used in This Study Antibody Source Catalogue number Application Working dilution DDIT4 Abcam ab106356 WB, IHC, IF WB 1: 1000 IHC 1: 600 IF 1: 100 mTOR Cell Signaling Technology #2983 WB 1: 1000 p-mTOR Cell Signaling Technology #5566 WB 1: 1000 ALOX15 Abcam ab244205 WB, IHC WB 1: 1000 IHC 1: 500 HMOX1 ZenBio R24541 WB, IHC WB 1: 1000 IHC 1: 400 GPX4 Proteintech #67763-1-1g WB, IHC WB 1: 1000 IHC 1: 1000 ATP4A Abcam ab231729 WB, IF, IHC WB 1: 1000 IF 1: 100 IHC 1: 200 ATP4B ZenBio #120298 WB, IHC WB 1:1000 IHC 1: 200 GIF Santacruz sc-514523 IF 1:50 GSII, Alexa FluorTM 647 Invitrogen L32451 IF 1:800 CD44V9 Novus Biologicals NBP2-53204 IF 1:100 MUC2 Cell Signaling Technology #88686 IF 1: 100 FABP1 Cell Signaling Technology #13368 IF 1:100 gH2AX Cell Signaling Technology 9718 IF 1: 100 TFF2 Thermo Fisher PA5-80111 WB 1: 1000 GAPDH ZenBio R24404 WB 1: 1000 b-actin ZenBio R51031 WB 1: 2000 IF, immunofluorescence; IHC, immunohistochemistry; WB, Western blot.

Techniques: Western Blot, Expressing, Immunohistochemistry, Staining, Microscopy, Infection, Activation Assay