phosphorylated p perk  (Cell Signaling Technology Inc)

Journal: Experimental and Therapeutic Medicine

Article Title: Peroxisome proliferator‑activated receptor γ alleviates human umbilical vein endothelial cell injury in deep vein thrombosis by blocking endoplasmic reticulum stress

doi: 10.3892/etm.2024.12674

Figure Lengend Snippet: PPARγ weakens the activation of endoplasmic reticulum stress in M-HSA-stimulated HUVECs. The protein expression levels of CHOP, GRP78, p-PERK, PERK, IRE1α and p-IRE1α were detected using western blot analysis. *** P<0.001 vs. HSA; # P<0.05, ## P<0.01 and ### P<0.001 vs. M-HSA + oe-NC. PPARγ, proliferator-activated receptor γ; M-HSA, modified glycated human serum albumin; HUVECs, human umbilical vein endothelial cells; CHOP, C/EBP homologous protein; GRP78, glucose-regulated protein 78; p, phosphorylated; PERK, protein kinase (PKR)-like ER kinase; IRE1α, p-inositol requiring enzyme 1α; oe, overexpressing; NC, negative control.

Article Snippet: Following blocking with 5% non-fat milk at room temperature for 2 h, the membranes were incubated with primary antibodies against PPARγ (1:1,000; cat. no. ab178860; Abcam), C/EBP homologous protein (CHOP; 1:1,000; cat. no. 2895; Cell Signaling Technology, Inc.), glucose-regulated protein 78 (GRP78; 1:1,000; cat. no. ab21685; Abcam), phosphorylated (p)-protein kinase (PKR)-like ER kinase (p-PERK; 1:200; cat. no. orb504147; Biorbyt), PERK (1:500; cat. no. orb1294328; Biorbyt), p-inositol requiring enzyme 1α (p-IRE1α; 1:1,000; cat. no. ab243665; Abcam), IRE1α (1:1,000; cat. no. ab37073; Abcam) and GAPDH (1:2,500; cat. no. ab9485; Abcam) at 4˚C overnight.

Techniques: Activation Assay, Expressing, Western Blot, Modification, Negative Control

Journal: Experimental Animals

Article Title: Daphnetin ameliorates diabetic cardiomyopathy by regulating inflammation and endoplasmic reticulum stress-induced apoptosis

doi: 10.1538/expanim.24-0027

Figure Lengend Snippet: Daphnetin suppresses endoplasmic reticulum (ER) stress-induced apoptosis in diabetic rats. (A) The expression levels of GRP78, CHOP, p-PERK, and PERK were examined by western blotting. The ratio of p-PERK/PERK was calculated. GAPDH was utilized as the internal control. (B) GRP78 and CHOP expression levels were quantified by immunofluorescence staining. (C) The number of apoptotic cells in myocardial tissues was determined by TUNEL assay. (D) The levels of Bax, Bcl-2, and cleaved caspase-3 in myocardial tissues were examined by western blotting. GAPDH was utilized as the internal control. ## indicates P <0.01 compared to the control group. * indicates P <0.05 and ** indicates P <0.01 compared to the STZ group.

Article Snippet: After blocking, the membranes were probed with antibodies against collagen I (#ab270993; 1:1,000; Abcam, Cambridge, MA, USA), collagen III (#ab184993; 1:1,000; Abcam), B cell lymphoma-2 (Bcl-2; #26593-1-AP; 1:3,000; Proteintech), Bcl-2-associated X protein (Bax; #2772; 1:1,000; CST, USA), cleaved caspase-3 (#9661; 1:1,000; CST, Shanghai, China), GRP78 (#11587-1-AP; 1:2,000; Proteintech), CHOP (#15204-1-AP; 1:1,500; Proteintech), protein kinase R-like ER kinase (PERK; #3192; 1:1,000; CST), phosphorylated (p)-PERK (#3179; 1:1,000; CST), c-Jun N-terminal kinase (JNK; #17572-1-AP; 1:3,000; Proteintech), p-JNK (#4668; 1:1,000; CST), p38 mitogen-activated protein kinase (p38 MAPK; #9212; 1:1,000; CST), p-p38 MAPK (#28796-1-AP; 1:1,000; Proteintech).

Techniques: Expressing, Western Blot, Control, Immunofluorescence, Staining, TUNEL Assay