phospho perk thr981  (Santa Cruz Biotechnology)

Journal: International Journal of Molecular Sciences

Article Title: Valdecoxib Protects against Cell Apoptosis Induced by Endoplasmic Reticulum Stress via the Inhibition of PERK-ATF4-CHOP Pathway in Experimental Glaucoma

doi: 10.3390/ijms232112983

Figure Lengend Snippet: Effect of valdecoxib on the expression of ER stress-related and apoptosis-related proteins in the OGD/R model. ( A ) Western blot analysis of p-PERK, ATF4, GRP78, CHOP, cleaved caspase 3, bax and bcl-2 levels in the control, OGD/R, OGD/R+DMSO and OGD/R+VAL groups. ( B – H ) Quantification of expression levels of p-PERK, ATF4, GRP78, CHOP, cleaved caspase 3, bax and bcl-2 in the control, OGD/R, OGD/R+DMSO and OGD/R+VAL groups using the densitometric analyses of Western blotting. The bar charts show the quantitative data (normalized by β-tubulin) for each protein relative to the control group (assigned a value of 1). Data are represented as the mean ± SD of three independent experiments. One-way ANOVA is used in B to H. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. control group. # p < 0.05, ## p < 0.01, ### p < 0.001 vs. OGD/R group. $ p < 0.05, $$ p < 0.01, $$$ p < 0.001 vs. OGD/R+DMSO group.

Article Snippet: We purchased propidium iodide (Cat: #81845, Sigma, St. Louis, MO, USA); Cell Counting Kit-8 (Cat: C0037, Beyotime, Shanghai, China), Annexin V-FITC/PI Kit (Cat: MA0220, Meilunbio, Dalian, China), TUNEL BrightRed Apoptosis Detection Kit (Cat: A113-03, Vazyme Biotech, Nanjing, China), agonist: CCT020312 (Cat: # HY-119240, MedChemExpress, Monmouth Junction, NJ, USA), primary antibody: RBPMS (Cat: 1830-RBPMS, PhosphoSolutions, Aurora, CO, USA, 1:200), GRP78 antibody (Cat: R24509, Zenbio, Chengdu, China, 1:1000), CHOP antibody (Cat: 1120880, Wanleibio, Shenyang, China, 1:1000), ATF4 (Cat: 381426, Zenbio, Chengdu, China, 1:500), PERK (phospho Thr981; Cat: YP1055, ImmunoWay Biotechnology Company, Plano, USA, 1:1000), bax (Cat: 50599-2-Ig, Proteintech, Wuhan, China, 1:2000), bcl-2 (Cat: 12789-1-AP, Proteintech, Wuhan, China, 1:1000) and caspase 3 (Cat: 19677-1-AP, Proteintech, Wuhan, China, 1:500).

Techniques: Expressing, Western Blot, Control

Journal: International Journal of Molecular Sciences

Article Title: Valdecoxib Protects against Cell Apoptosis Induced by Endoplasmic Reticulum Stress via the Inhibition of PERK-ATF4-CHOP Pathway in Experimental Glaucoma

doi: 10.3390/ijms232112983

Figure Lengend Snippet: Effect of valdecoxib on the expression of ER stress-related and apoptosis-related proteins in the retinal ischemia-reperfusion model. ( A ) Western blot analysis of p-PERK, ATF4, GRP78, CHOP, cleaved caspase 3, bax and bcl-2 levels in the control, I/R, I/R+DMSO and I/R+VAL groups. β-tubulin served as the loading control. ( B – H ) Quantification of expression levels of p-PERK, ATF4, GRP78, CHOP, cleaved caspase 3, bax and bcl-2 in the control, I/R, I/R+DMSO and I/R+VAL groups using the densitometric analyses of Western blotting. The bar charts show the quantitative data (normalized by β-tubulin) for each protein relative to the sham group (assigned a value of 1). ( I ) Representative fluorescence images of p-PERK staining are shown (scale bar = 50 μm). Immunostaining was executed using a primary antibody against p-PERK (green), and the nucleus (blue) is marked by DAPI. ( J ) Representative fluorescence images of cleaved caspase 3 staining are shown (scale bar = 50 μm). Immunostaining was executed using a primary antibody against cleaved caspase 3 (green), and the nucleus (blue) is marked by DAPI. ( K ) Representative fluorescence images of GRP78 staining are shown (scale bar = 50 μm). Immunostaining was executed using a primary antibody against GRP78 (green), and the nucleus (blue) is marked by DAPI. Images demonstrate the increased expression level of p-PERK, cleaved caspase 3 and GRP78 in the I/R and I/R+DMSO groups, and decreased expression level of those proteins in the I/R+VAL group in the retinal ganglion cell layer. Data are represented as the mean ± SD of three independent experiments. Each group was composed of five rats. One-way ANOVA is used in B to H. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. sham group. ## p < 0.01, ### p < 0.001 vs. I/R group. $$ p < 0.01, $$$ p < 0.001 vs. I/R+DMSO group.

Article Snippet: We purchased propidium iodide (Cat: #81845, Sigma, St. Louis, MO, USA); Cell Counting Kit-8 (Cat: C0037, Beyotime, Shanghai, China), Annexin V-FITC/PI Kit (Cat: MA0220, Meilunbio, Dalian, China), TUNEL BrightRed Apoptosis Detection Kit (Cat: A113-03, Vazyme Biotech, Nanjing, China), agonist: CCT020312 (Cat: # HY-119240, MedChemExpress, Monmouth Junction, NJ, USA), primary antibody: RBPMS (Cat: 1830-RBPMS, PhosphoSolutions, Aurora, CO, USA, 1:200), GRP78 antibody (Cat: R24509, Zenbio, Chengdu, China, 1:1000), CHOP antibody (Cat: 1120880, Wanleibio, Shenyang, China, 1:1000), ATF4 (Cat: 381426, Zenbio, Chengdu, China, 1:500), PERK (phospho Thr981; Cat: YP1055, ImmunoWay Biotechnology Company, Plano, USA, 1:1000), bax (Cat: 50599-2-Ig, Proteintech, Wuhan, China, 1:2000), bcl-2 (Cat: 12789-1-AP, Proteintech, Wuhan, China, 1:1000) and caspase 3 (Cat: 19677-1-AP, Proteintech, Wuhan, China, 1:500).

Techniques: Expressing, Western Blot, Control, Fluorescence, Staining, Immunostaining

Journal: International Journal of Molecular Sciences

Article Title: Valdecoxib Protects against Cell Apoptosis Induced by Endoplasmic Reticulum Stress via the Inhibition of PERK-ATF4-CHOP Pathway in Experimental Glaucoma

doi: 10.3390/ijms232112983

Figure Lengend Snippet: PERK activator CCT020312 reverses the effect of valdecoxib on the expression of ER stress-related proteins and on the apoptosis-related proteins in the OGD/R model. R28 cells were pretreated with CCT020312 at the concentration of 5 μmol/L for 4 h. ( A ) Western blot analysis of p-PERK, ATF4, GRP78, CHOP, cleaved caspase 3, bax and bcl-2 levels in the control, OGD/R+DMSO, OGD/R+VAL, OGD/R+VAL+CCT and OGD/R+DMSO+CCT groups. β-tubulin served as the loading control. ( B – H ) Quantification of expression levels of p-PERK, ATF4, GRP78, CHOP, cleaved caspase 3, bax and bcl-2 in the control, OGD/R+DMSO, OGD/R+VAL, OGD/R+VAL+CCT and OGD/R+DMSO+CCT groups using the densitometric analyses of Western blotting. The bar charts show the quantitative data (normalized by β-tubulin) for each protein relative to the control group (assigned a value of 1). Data are represented as the mean ± SD of three independent experiments. One-way ANOVA is used in B to H. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. control group. # p < 0.05, ## p < 0.01, ### p < 0.001 vs. OGD/R+DMSO group. $ p < 0.05, $$ p < 0.01, $$$ p < 0.001 vs. OGD/R+VAL group.

Article Snippet: We purchased propidium iodide (Cat: #81845, Sigma, St. Louis, MO, USA); Cell Counting Kit-8 (Cat: C0037, Beyotime, Shanghai, China), Annexin V-FITC/PI Kit (Cat: MA0220, Meilunbio, Dalian, China), TUNEL BrightRed Apoptosis Detection Kit (Cat: A113-03, Vazyme Biotech, Nanjing, China), agonist: CCT020312 (Cat: # HY-119240, MedChemExpress, Monmouth Junction, NJ, USA), primary antibody: RBPMS (Cat: 1830-RBPMS, PhosphoSolutions, Aurora, CO, USA, 1:200), GRP78 antibody (Cat: R24509, Zenbio, Chengdu, China, 1:1000), CHOP antibody (Cat: 1120880, Wanleibio, Shenyang, China, 1:1000), ATF4 (Cat: 381426, Zenbio, Chengdu, China, 1:500), PERK (phospho Thr981; Cat: YP1055, ImmunoWay Biotechnology Company, Plano, USA, 1:1000), bax (Cat: 50599-2-Ig, Proteintech, Wuhan, China, 1:2000), bcl-2 (Cat: 12789-1-AP, Proteintech, Wuhan, China, 1:1000) and caspase 3 (Cat: 19677-1-AP, Proteintech, Wuhan, China, 1:500).

Techniques: Expressing, Concentration Assay, Western Blot, Control

Journal: eLife

Article Title: Impaired mRNA splicing and proteostasis in preadipocytes in obesity-related metabolic disease

doi: 10.7554/eLife.65996

Figure Lengend Snippet:

Article Snippet: Antibody , Phospho-PERK (Thr981)(rabbit polyclonal) , Santa Cruz Biotechnology , sc-32577, AB_2293243 , (1:750).

Techniques: Ubiquitin Proteomics, Recombinant, Sequencing, Negative Control, Western Blot, Saline, Modification, Fluorescence, Multiplex sample analysis, Transfection, Red Blood Cell Lysis, Mass Spectrometry, Binding Assay, DC Protein Assay, cDNA Synthesis, Software, High Content Screening, Cell Analysis, Real-time Polymerase Chain Reaction