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phosphorylated eif 2α  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phosphorylated eif 2α
    Phosphorylated Eif 2α, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1405 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/eif+2%CE%B1/Phospho-eIF2alpha+(Ser51)+XP+Rabbit+mAb/pmc12907968-8-0-7
    Average 96 stars, based on 1405 article reviews
    phosphorylated eif 2α - by Bioz Stars, 2026-09
    96/100 stars

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    Related Articles

    Blocking Assay:

    Article Title: GPX1-driven selenium nanoplatform reprograms MAMs-mediated organelle crosstalk to reverse inflammatory adipose expansion in thyroid eye disease
    Article Snippet: Equal protein amounts were resolved by SDS-PAGE and transferred onto PVDF membranes (Millipore, IPVH00010). .. After blocking with 5% (w/v) skimmed milk (1 h), membranes were incubated overnight at 4 °C with primary antibodies against GPX1 (Abcam, ab22604), β-actin (Cell Signaling Technology [CST], Danvers, MA, USA; #4967), FABP4 (CST, #2120), CEBPA (Abcam, ab317442), PLIN1 (Abcam, ab172907), PPARG (CST, #2435), BiP (CST, #3183), PERK (CST, #3192), p-PERK (Thr980; Invitrogen, MA5-15033), eIF-2α (CST, #9722), p-eIF-2α (Ser51; CST, #9721), ATF4 (CST, #11815), CHOP (CST, #5554), GRP75 (CST, #2816), VDAC1 (CST, #4866), IP3R1 (CST, #8568), STING (Proteintech, 19851-1-AP), p-STING (Ser365; CST, #72971), TBK1 (CST, #3504), pTBK1 (Ser172; CST, #5483), IRF3 (CST, #4302), p-IRF3 (Ser396; CST, #29047), NF-κB (CST, #8242), and p-NF-κB (Ser536; CST, #3033). .. HRP-conjugated secondary antibodies (CST, #7074S) were applied (1 h, room temperature), and protein bands were visualized using an ECL reagent (Thermo Fisher, 34577).

    Incubation:

    Article Title: GPX1-driven selenium nanoplatform reprograms MAMs-mediated organelle crosstalk to reverse inflammatory adipose expansion in thyroid eye disease
    Article Snippet: Equal protein amounts were resolved by SDS-PAGE and transferred onto PVDF membranes (Millipore, IPVH00010). .. After blocking with 5% (w/v) skimmed milk (1 h), membranes were incubated overnight at 4 °C with primary antibodies against GPX1 (Abcam, ab22604), β-actin (Cell Signaling Technology [CST], Danvers, MA, USA; #4967), FABP4 (CST, #2120), CEBPA (Abcam, ab317442), PLIN1 (Abcam, ab172907), PPARG (CST, #2435), BiP (CST, #3183), PERK (CST, #3192), p-PERK (Thr980; Invitrogen, MA5-15033), eIF-2α (CST, #9722), p-eIF-2α (Ser51; CST, #9721), ATF4 (CST, #11815), CHOP (CST, #5554), GRP75 (CST, #2816), VDAC1 (CST, #4866), IP3R1 (CST, #8568), STING (Proteintech, 19851-1-AP), p-STING (Ser365; CST, #72971), TBK1 (CST, #3504), pTBK1 (Ser172; CST, #5483), IRF3 (CST, #4302), p-IRF3 (Ser396; CST, #29047), NF-κB (CST, #8242), and p-NF-κB (Ser536; CST, #3033). .. HRP-conjugated secondary antibodies (CST, #7074S) were applied (1 h, room temperature), and protein bands were visualized using an ECL reagent (Thermo Fisher, 34577).

    Article Title: Factors affecting the expression and stability of full-length and truncated SRSF3 proteins in human cancer cells
    Article Snippet: The protein lysate was then prepared with 4 × protein loading dye, denatured at 95 °C for 10 min, separated via 12% SDS‒PAGE, and blotted onto PVDF membranes (Immobilon-P; Millipore, Bedford, MA, USA) using a Bio-Rad Semi-Dry Transfer Cell. .. The blots were then incubated with primary antibodies against α-actinin (ACTN, H-2, sc-17829), p53 (DO-1, sc-125), c-Jun (H-79, sc-1694), XBP-1 (F-4, sc-8015), and C/EBPβ (H-7, sc-7962) (Santa Cruz Biotechnology, USA); cleaved PARP (#9546), p-eIF2α (#9721), eIF-2α (#9722), Upf-1 (#12,040), and HIF-1α (#14,179) (Cell Signaling, Danvers, MA, USA); cyclin D1 (ab134175), p21 (ab109520), and γH2AX (ab81299) (Abcam, Cambridge, UK); SRSF3 (WH0006428M8, Sigma‒Aldrich); and HA (clone 3F10, Roche). .. The blots were subsequently incubated with HRP-conjugated secondary antibodies (anti-mouse IgG, AP192P; and anti-rabbit IgG, AP132P; Merck-Millipore).

    Produced:

    Article Title: IRE1α modulates M1 oncolytic virus sensitivity via ER stress regulation in bladder cancer
    Article Snippet: .. Antibodies were listed as following: IRE1α (1:1,000, ab96481, Abcam, Cambridge, UK), PERK (1:1,000, #5683, Cell Signaling Technology, Irving, TX, USA), ATF6α (1:1,000, #65880, Cell Signaling Technology, Irving, TX, USA), eIF-2α (1:1,000, #5324, Cell Signaling Technology, Irving, TX, USA), phosphorylated eIF-2α (1:1,000, #3398, Cell Signaling Technology, Irving, TX, USA), GAPDH (1:1,000, AP0060, Bioworld), β-actin (1:1,000, AP0063, Bioworld), JNK (1:1,000, #9252, Cell Signaling Technology, Irving, TX, USA), phosphorylated JNK (1:1,000, #925, Cell Signaling Technology, Irving, TX, USA), M1 E1 and NS3 (1:1,000, produced by Beijing Protein Innovation), CHOP (1:1,000, ab11419, Abcam, Cambridge, UK), XBP1 (1:1,000, #40435, Cell Signaling Technology, Irving, TX, USA), ZAP (1:1,000, #PA5-31650, Thermo Scientific, USA), Caspase-12 (1:1,000, #58208, Cell Signaling Technology, Irving, TX, USA) and cleaved-Caspase-3 (1:1,000, #9664, Cell Signaling Technology, Irving, TX, USA). ..

    Article Title: RIPK3 deficiency ameliorates diabetic sarcopenia through proteostasis regulation by suppressing inflammatory signaling and cellular stress pathways.
    Article Snippet: Diabetic sarcopenia is a major complication of diabetes severely impacting patient quality of life and increasing mortality risk.. Its pathogenesis remains incompletely understood and effective treatments are lacking.. This study reveals the critical role and molecular mechanisms of Receptor-interacting protein kinase 3 (RIPK3) in skeletal muscle atrophy during type 1 diabetes mellitus (T1DM).

    Membrane:

    Article Title: Overexpression of mitofusin 2 ameliorates inflammation and oxidative stress in lipopolysaccharide-induced mastitis model by regulating phosphofurin acidic cluster sorting protein 2.
    Article Snippet: .. Antibody name Catalog number Dilution factor Company Cyclooxygenase- 2 27308- 1- AP 1:1500 Proteintech Inducible nitric oxide synthase 22226- 1- AP 1:1000 Proteintech Mitochondrial fusion protein 2 12186- 1- AP 1:2500 Proteintech Mitochondrial fusion protein 1 13798- 1- AP 1:2500 Proteintech Mitochondrial fission protein 1 10956- 1- AP 1:4000 Proteintech Dynamin- related protein 1 8570 1:2000 Cell Signaling Technology Optic atrophy 1 80471 1:2000 Cell Signaling Technology Eukaryotic translation initiation factor 2 alpha (eIF- 2α) 9722 1:1000 Cell Signaling Technology Phosphorylated- eIF- 2α (P- eIF- 2α) 3398 1:1000 Cell Signaling Technology Phosphacinosin- membrane- associated protein 2 30172- 1- AP 1:1000 Proteintech β- Actin 66009- 1- Ig 1:5000 Proteintech TA B L E 2 Primer sequences. ..



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    Image Search Results


    HSV-1 neurotropic infection causes mitochondrial damage in vivo and in vitro . ( A ) Distribution of viral gene in different brain region of HSE mice (n = 6). (B) Immunohistochemistry assay was performed to detect mitochondrial TOMM20 in various brain regions, including olfactory bulb (OB), cerebral cortex (CC), and pons/medulla oblongata/cerebellum (P/M/C) of both mock-infected (Ctrl) and HSV-1 infected mice. Scale bar, 100 μm. (C) TEM analysis of mitochondria in the brain tissue from control or HSE mice. ( D ) Quantification of damaged and total mitochondria from 15 fields of view, as well as mitochondrial aspect ratio and form factor. ( E ) Western blot assay of mitophagy-related proteins the brain tissue of control or HSE mice (n = 3). ( F ) RT-qPCR analysis of the mRNA expression levels of PRKN and PINK1 in OB or total brain tissues. ( G ) BV2 and N2a cells were infected with HSV-1 (MOI 0.1, 0.5 and 1) or treated with CCCP (2 μM) for 12 h, followed by staining with JC-1 probe. Flow cytometry was employed to measure the mitochondrial membrane potential. (H) BV2 cells were infected with HSV-1 (MOI=1) for indicated hours and flow cytometry assay was performed to detect the mitochondrial membrane potential. (I) BV2 cells were infected with HSV-1 (MOI=1) for 12 h and RT-qPCR assay was used to detect the relative mRNA expression ratio of the mitochondrial gene mtCO1 and the nuclear gene β-globin . ( J ) Flow cytometry analysis of intracellular ROS production. ( K ) Western blot analysis of proteins involved in mitochondrial fusion or fission. Data are means ± SD (n = 3). * p < 0.05, ** p < 0.01 or *** p < 0.001 versus control group.

    Journal: Journal of Advanced Research

    Article Title: Inhibition of mitophagy via the EIF2S1-ATF4-PRKN pathway contributes to viral encephalitis

    doi: 10.1016/j.jare.2024.08.003

    Figure Lengend Snippet: HSV-1 neurotropic infection causes mitochondrial damage in vivo and in vitro . ( A ) Distribution of viral gene in different brain region of HSE mice (n = 6). (B) Immunohistochemistry assay was performed to detect mitochondrial TOMM20 in various brain regions, including olfactory bulb (OB), cerebral cortex (CC), and pons/medulla oblongata/cerebellum (P/M/C) of both mock-infected (Ctrl) and HSV-1 infected mice. Scale bar, 100 μm. (C) TEM analysis of mitochondria in the brain tissue from control or HSE mice. ( D ) Quantification of damaged and total mitochondria from 15 fields of view, as well as mitochondrial aspect ratio and form factor. ( E ) Western blot assay of mitophagy-related proteins the brain tissue of control or HSE mice (n = 3). ( F ) RT-qPCR analysis of the mRNA expression levels of PRKN and PINK1 in OB or total brain tissues. ( G ) BV2 and N2a cells were infected with HSV-1 (MOI 0.1, 0.5 and 1) or treated with CCCP (2 μM) for 12 h, followed by staining with JC-1 probe. Flow cytometry was employed to measure the mitochondrial membrane potential. (H) BV2 cells were infected with HSV-1 (MOI=1) for indicated hours and flow cytometry assay was performed to detect the mitochondrial membrane potential. (I) BV2 cells were infected with HSV-1 (MOI=1) for 12 h and RT-qPCR assay was used to detect the relative mRNA expression ratio of the mitochondrial gene mtCO1 and the nuclear gene β-globin . ( J ) Flow cytometry analysis of intracellular ROS production. ( K ) Western blot analysis of proteins involved in mitochondrial fusion or fission. Data are means ± SD (n = 3). * p < 0.05, ** p < 0.01 or *** p < 0.001 versus control group.

    Article Snippet: Antibodies against HSV-1 gB (sc-56987), eIF-2α (sc-133132), phospho-eIF-2α (sc-310073), DRP1 (sc-271583), OPA1 (sc-393296), MFN1 (sc-166644) and MFN2 (sc-100560) were purchased from Santa Cruz Biotechnology (California, USA).

    Techniques: Infection, In Vivo, In Vitro, Immunohistochemistry, Control, Western Blot, Quantitative RT-PCR, Expressing, Staining, Flow Cytometry, Membrane

    HSV-1 inhibits mitophagy. ( A ) BV2 and N2a cells were infected with HSV-1 (MOI=1) for indicated times and the protein expression levels of PINK1, PRKN and TOMM20 were analyzed by Western blot. (B-C) Immunoblot analysis of indicated proteins in BV2 cells infected with HSV-1 for 3 h at various MOIs. ( C ) Immunoblot analysis of indicated proteins in BV2 cells infected with HSV-1 in the presence or absence of CCCP (2 μM) for indicated times. ( D ) BV2 cells were infected with HSV-1 for indicated times and the mRNA expression levels of PINK1 and PRKN were analyzed by RT-qPCR assay. Data are means ± SD (n = 3). * p < 0.05, ** p < 0.01 versus control group. ( E ) TEM analysis of mitochondria in BV2 cells infected with HSV-1 for 24 h. Arrows indicate viral particles. Scale bar, 2 μm. ( F ) Quantification of damaged mitochondria from 10 fields of view, as well as mitochondrial aspect ratio and form factor. * p < 0.05, ** p < 0.01, *** p < 0.001versus control group.

    Journal: Journal of Advanced Research

    Article Title: Inhibition of mitophagy via the EIF2S1-ATF4-PRKN pathway contributes to viral encephalitis

    doi: 10.1016/j.jare.2024.08.003

    Figure Lengend Snippet: HSV-1 inhibits mitophagy. ( A ) BV2 and N2a cells were infected with HSV-1 (MOI=1) for indicated times and the protein expression levels of PINK1, PRKN and TOMM20 were analyzed by Western blot. (B-C) Immunoblot analysis of indicated proteins in BV2 cells infected with HSV-1 for 3 h at various MOIs. ( C ) Immunoblot analysis of indicated proteins in BV2 cells infected with HSV-1 in the presence or absence of CCCP (2 μM) for indicated times. ( D ) BV2 cells were infected with HSV-1 for indicated times and the mRNA expression levels of PINK1 and PRKN were analyzed by RT-qPCR assay. Data are means ± SD (n = 3). * p < 0.05, ** p < 0.01 versus control group. ( E ) TEM analysis of mitochondria in BV2 cells infected with HSV-1 for 24 h. Arrows indicate viral particles. Scale bar, 2 μm. ( F ) Quantification of damaged mitochondria from 10 fields of view, as well as mitochondrial aspect ratio and form factor. * p < 0.05, ** p < 0.01, *** p < 0.001versus control group.

    Article Snippet: Antibodies against HSV-1 gB (sc-56987), eIF-2α (sc-133132), phospho-eIF-2α (sc-310073), DRP1 (sc-271583), OPA1 (sc-393296), MFN1 (sc-166644) and MFN2 (sc-100560) were purchased from Santa Cruz Biotechnology (California, USA).

    Techniques: Infection, Expressing, Western Blot, Quantitative RT-PCR, Control

    HSV-1 deregulates the EIF2S1-ATF4 to suppress PINK1/PRKN expression. ( A ) Western blot analysis of the impact of HSV-1 infection on ATF4 protein levels. ( B ) CCCP (2 μM) treatment affects the protein levels of ATF4 in BV2 cells infected with HSV-1 for 3 h and 24 h, respectively. ( C-D ) Overexpression of ATF4 led to changes in the protein levels (C) and mRNA expression levels (D) of PINK1 and PRKN. Data are means ± SD (n = 3). * p < 0.05, ** p < 0.01 versus control group. ( E ) The effect of ATF4 knockdown on CCCP-mediated mitophagy. Cells were transfected with ATF4 siRNA for 48 h, followed by treatment with CCCP for 12 h. Total protein was subjected to western blot assay. ( F ) The effect of ATF4 knockdown on the mRNA expression of PINK1 and PRKN. Cells were transfected with ATF4 siRNA for 48 h and RT-qPCR assay was performed. Data are means ± SD (n = 3). * p < 0.05, ** p < 0.01 versus control group. ( G ) Western blot analysis of p-EIF2S1 and EIF2S1 in HSV-1 infected cells.

    Journal: Journal of Advanced Research

    Article Title: Inhibition of mitophagy via the EIF2S1-ATF4-PRKN pathway contributes to viral encephalitis

    doi: 10.1016/j.jare.2024.08.003

    Figure Lengend Snippet: HSV-1 deregulates the EIF2S1-ATF4 to suppress PINK1/PRKN expression. ( A ) Western blot analysis of the impact of HSV-1 infection on ATF4 protein levels. ( B ) CCCP (2 μM) treatment affects the protein levels of ATF4 in BV2 cells infected with HSV-1 for 3 h and 24 h, respectively. ( C-D ) Overexpression of ATF4 led to changes in the protein levels (C) and mRNA expression levels (D) of PINK1 and PRKN. Data are means ± SD (n = 3). * p < 0.05, ** p < 0.01 versus control group. ( E ) The effect of ATF4 knockdown on CCCP-mediated mitophagy. Cells were transfected with ATF4 siRNA for 48 h, followed by treatment with CCCP for 12 h. Total protein was subjected to western blot assay. ( F ) The effect of ATF4 knockdown on the mRNA expression of PINK1 and PRKN. Cells were transfected with ATF4 siRNA for 48 h and RT-qPCR assay was performed. Data are means ± SD (n = 3). * p < 0.05, ** p < 0.01 versus control group. ( G ) Western blot analysis of p-EIF2S1 and EIF2S1 in HSV-1 infected cells.

    Article Snippet: Antibodies against HSV-1 gB (sc-56987), eIF-2α (sc-133132), phospho-eIF-2α (sc-310073), DRP1 (sc-271583), OPA1 (sc-393296), MFN1 (sc-166644) and MFN2 (sc-100560) were purchased from Santa Cruz Biotechnology (California, USA).

    Techniques: Expressing, Western Blot, Infection, Over Expression, Control, Knockdown, Transfection, Quantitative RT-PCR

    Activation of mitophagy inhibits HSV-1 infection. ( A ) BV2 cells were infected with HSV-1 (MOI=0.1) or ACV-resistant strains HSV-1/Blue and HSV-1/153 (MOI=0.1) for 24 h in the presence of CCCP (2 μM), Rotenone (1 μM) or ACV (1 or 2 μM), and virus proliferation was assessed by plaque assay. ( B ) RT-qPCR analysis of the copy number of viral genes UL54 , U52 and UL27 in HSV-1-infected BV2 cells treated with CCCP or Rotenone. ( C-D ) Western blot analysis of mitophagy-related protein in cells that were transfected with PRKN -expressing plasmids for 48 h and subsequently infected with HSV-1 for 24 h. ( E-F ) The overexpression of PRKN inhibits viral plaque numbers (E) and the DNA copy number of viral genes (F). Data are means ± SD (n = 3). * p < 0.05, ** p < 0.01 or *** p < 0.001 versus control group. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

    Journal: Journal of Advanced Research

    Article Title: Inhibition of mitophagy via the EIF2S1-ATF4-PRKN pathway contributes to viral encephalitis

    doi: 10.1016/j.jare.2024.08.003

    Figure Lengend Snippet: Activation of mitophagy inhibits HSV-1 infection. ( A ) BV2 cells were infected with HSV-1 (MOI=0.1) or ACV-resistant strains HSV-1/Blue and HSV-1/153 (MOI=0.1) for 24 h in the presence of CCCP (2 μM), Rotenone (1 μM) or ACV (1 or 2 μM), and virus proliferation was assessed by plaque assay. ( B ) RT-qPCR analysis of the copy number of viral genes UL54 , U52 and UL27 in HSV-1-infected BV2 cells treated with CCCP or Rotenone. ( C-D ) Western blot analysis of mitophagy-related protein in cells that were transfected with PRKN -expressing plasmids for 48 h and subsequently infected with HSV-1 for 24 h. ( E-F ) The overexpression of PRKN inhibits viral plaque numbers (E) and the DNA copy number of viral genes (F). Data are means ± SD (n = 3). * p < 0.05, ** p < 0.01 or *** p < 0.001 versus control group. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

    Article Snippet: Antibodies against HSV-1 gB (sc-56987), eIF-2α (sc-133132), phospho-eIF-2α (sc-310073), DRP1 (sc-271583), OPA1 (sc-393296), MFN1 (sc-166644) and MFN2 (sc-100560) were purchased from Santa Cruz Biotechnology (California, USA).

    Techniques: Activation Assay, Infection, Virus, Plaque Assay, Quantitative RT-PCR, Western Blot, Transfection, Expressing, Over Expression, Control

    PRKN-overexpressing mice are resistant to HSV-1 infection and HSE. ( A ) Schematic diagram illustrating the administration process for LV-PRKN mice. Lentivirus (puro)-CMV-NC and Lentivirus (puro)-CMV-PRKN were nasally administered twice with an interval of 48 h before being inoculated intranasally with HSV-1 (2 × 10 6 PFU) to establish the HSE model (n = 12). ( B ) Western blot analysis of PRKN expression in OB tissue from LV-PRKN or LV-Ctrl HSE mice (n = 3). ( C-E ) The mice were daily scored for HSE-associated symptoms (C), eye damage (D), and survival rate (E). Data are mean ± SD (n = 12). ( F ) RT-qPCR analysis of IL-1β , IL-6 and TNF-α mRNA expression in different brain tissues, including cerebral cortex (CX), olfactory bulb (OB) and pons/medulla oblongata/cerebellum (P/M/C), at 8 days post HSV-1 infection (n = 5–6). * p < 0.05, ** p < 0.01 or *** p < 0.001 versus LV-Ctrl group. ( G ) Representative images of H&E staining of different brain tissues from HSE mice (n = 3). Scale bar, 100 μm. ( H ) Immunohistochemistry analysis was performed to detect the viral load (anti-VP5) in the CX region (n = 3). Scale bar, 100 μm. ( I ) The HSV-1 DNA copy number in the brain of LV-Ctrl and LV-PRKN group was examined by RT-qPCR assay at 8 d.p.i (n = 5).

    Journal: Journal of Advanced Research

    Article Title: Inhibition of mitophagy via the EIF2S1-ATF4-PRKN pathway contributes to viral encephalitis

    doi: 10.1016/j.jare.2024.08.003

    Figure Lengend Snippet: PRKN-overexpressing mice are resistant to HSV-1 infection and HSE. ( A ) Schematic diagram illustrating the administration process for LV-PRKN mice. Lentivirus (puro)-CMV-NC and Lentivirus (puro)-CMV-PRKN were nasally administered twice with an interval of 48 h before being inoculated intranasally with HSV-1 (2 × 10 6 PFU) to establish the HSE model (n = 12). ( B ) Western blot analysis of PRKN expression in OB tissue from LV-PRKN or LV-Ctrl HSE mice (n = 3). ( C-E ) The mice were daily scored for HSE-associated symptoms (C), eye damage (D), and survival rate (E). Data are mean ± SD (n = 12). ( F ) RT-qPCR analysis of IL-1β , IL-6 and TNF-α mRNA expression in different brain tissues, including cerebral cortex (CX), olfactory bulb (OB) and pons/medulla oblongata/cerebellum (P/M/C), at 8 days post HSV-1 infection (n = 5–6). * p < 0.05, ** p < 0.01 or *** p < 0.001 versus LV-Ctrl group. ( G ) Representative images of H&E staining of different brain tissues from HSE mice (n = 3). Scale bar, 100 μm. ( H ) Immunohistochemistry analysis was performed to detect the viral load (anti-VP5) in the CX region (n = 3). Scale bar, 100 μm. ( I ) The HSV-1 DNA copy number in the brain of LV-Ctrl and LV-PRKN group was examined by RT-qPCR assay at 8 d.p.i (n = 5).

    Article Snippet: Antibodies against HSV-1 gB (sc-56987), eIF-2α (sc-133132), phospho-eIF-2α (sc-310073), DRP1 (sc-271583), OPA1 (sc-393296), MFN1 (sc-166644) and MFN2 (sc-100560) were purchased from Santa Cruz Biotechnology (California, USA).

    Techniques: Infection, Western Blot, Expressing, Quantitative RT-PCR, Staining, Immunohistochemistry