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p ire1α  (Novus Biologicals)


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    Structured Review

    Novus Biologicals p ire1α
    P Ire1α, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 96/100, based on 243 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/p+ire1%CE%B1/IRE1+alpha+%5Bp+Ser724%5D+Antibody+-+BSA+Free/us12605424-306-40-42
    Average 96 stars, based on 243 article reviews
    p ire1α - by Bioz Stars, 2026-11
    96/100 stars

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    Related Articles

    Incubation:

    Article Title: The PERK-eIF2α-ATF4 Axis Is Involved in Mediating ER-Stress-Induced Ferroptosis via DDIT4-mTORC1 Inhibition and Acetaminophen-Induced Hepatotoxicity.
    Article Snippet: Samples were boiled at 95 ◦C in 2x×Laemmli Sample Buffer (1610737, Bio-Rad, Hercules, CA, USA) for 5 min. Proteins were resolved by SDS-PAGE and transferred to polyvinylidene difluoride membranes (IPVH00010, Millipore, Burlington, MA, USA). .. The membranes were blocked with 5% nonfat milk (BD232100, BD bioscience, San Diego, CA, USA) in phosphate-buffered salineTween 20 (PBS-T), and then the membranes were incubated with primary antibodies as follows: p-PERK (1:1000, Cat#12814, Signalway antibody, Greenbelt, MD, USA), PERK (1:1000, Cat#5683, Cell Signaling, Danvers, MA, USA), p-eIF2α (1:1000, Cat#9721, Cell Signaling, Danvers, MA, USA), eIF2α (1:1000, Cat#9722, Cell Signaling, Danvers, MA, USA), ATF4 (1:1000, Cat#10835-1-AP, Proteintech, Rosemont, IL, USA), GRP78 (1:1000, Cat#ADI-SPA-826, Enzo Life Sciences, Farmingdale, NY, USA), DDIT3 (1:500, Cat#sc7351, Santa Cruz, Dallas, TX, USA), p-IRE1α (1:1000, Cat#NB100-2323, Novus Biologicals, Centennial, CO, USA), total-IRE1α (1:500, Cat#sc-390960, Santa Cruz, Dallas, TX, USA), ATF6 (1:1000, #Cat24169-AP, Proteintech, Rosemont, IL, USA), XBP-1(s) (1:1000, Cat#647502, BioLegend, San Diego, CA, USA), GPX4 (1:2000, Cat#Ab125066, Abcam, Cambridge, UK), SLC7A11 (1:1000, Cat#PA1-16775, Thermo Fisher Scientific, Waltham, MA, USA), MDA (1:1000, Cat#6463, Abcam, Cambridge, UK), 4-HNE (1:1000, Cat#46545, Abcam, Cambridge, UK), CHAC1 (1:1000, Cat#15207-AP, Proteintech, Rosemont, IL, USA), DDIT4 (1:1000, Cat#10638-1-AP, Proteintech, Rosemont, IL, USA), cleaved caspase-3 (1:1000, Cat#9661, Cell signaling, Danvers, MA, USA), caspase-3 (1:1000, Cat#9662, Cell signaling, Danvers, MA, USA), p-mTOR (1:1000, Cat#2971, Cell signaling, Danvers, MA, USA), total-mTOR (1:1000, Cat#2972, Cell signaling, Danvers, MA, USA), p-4EBP1 (1:1000, Cat#9455, Cell signaling, Danvers, MA, USA), total-4EBP1 (1:1000, Cat#9452, Cell signaling, Danvers, MA, USA), p-S6 ribosomal protein (1:2000, Cat#5364, Cell signaling, Danvers, MA, USA), S6 ribosomal protein (1:2000, Cat#2317, Cell signaling, Danvers, MA, USA), and β-actin (1:2000, #Cat4967, Cell signaling, Danvers, MA, USA) as a loading control. ..

    Article Title: The PERK-eIF2α-ATF4 Axis Is Involved in Mediating ER-Stress-Induced Ferroptosis via DDIT4-mTORC1 Inhibition and Acetaminophen-Induced Hepatotoxicity
    Article Snippet: Samples were boiled at 95 °C in 2x×Laemmli Sample Buffer (1610737, Bio-Rad, Hercules, CA, USA) for 5 min. Proteins were resolved by SDS-PAGE and transferred to polyvinylidene difluoride membranes (IPVH00010, Millipore, Burlington, MA, USA). .. The membranes were blocked with 5% nonfat milk (BD232100, BD bioscience, San Diego, CA, USA) in phosphate-buffered saline-Tween 20 (PBS-T), and then the membranes were incubated with primary antibodies as follows: p-PERK (1:1000, Cat#12814, Signalway antibody, Greenbelt, MD, USA), PERK (1:1000, Cat#5683, Cell Signaling, Danvers, MA, USA), p-eIF2α (1:1000, Cat#9721, Cell Signaling, Danvers, MA, USA), eIF2α (1:1000, Cat#9722, Cell Signaling, Danvers, MA, USA), ATF4 (1:1000, Cat#10835-1-AP, Proteintech, Rosemont, IL, USA), GRP78 (1:1000, Cat#ADI-SPA-826, Enzo Life Sciences, Farmingdale, NY, USA), DDIT3 (1:500, Cat#sc-7351, Santa Cruz, Dallas, TX, USA), p-IRE1α (1:1000, Cat#NB100-2323, Novus Biologicals, Centennial, CO, USA), total-IRE1α (1:500, Cat#sc-390960, Santa Cruz, Dallas, TX, USA), ATF6 (1:1000, #Cat24169-AP, Proteintech, Rosemont, IL, USA), XBP-1(s) (1:1000, Cat#647502, BioLegend, San Diego, CA, USA), GPX4 (1:2000, Cat#Ab125066, Abcam, Cambridge, UK), SLC7A11 (1:1000, Cat#PA1-16775, Thermo Fisher Scientific, Waltham, MA, USA), MDA (1:1000, Cat#6463, Abcam, Cambridge, UK), 4-HNE (1:1000, Cat#46545, Abcam, Cambridge, UK), CHAC1 (1:1000, Cat#15207-AP, Proteintech, Rosemont, IL, USA), DDIT4 (1:1000, Cat#10638-1-AP, Proteintech, Rosemont, IL, USA), cleaved caspase-3 (1:1000, Cat#9661, Cell signaling, Danvers, MA, USA), caspase-3 (1:1000, Cat#9662, Cell signaling, Danvers, MA, USA), p-mTOR (1:1000, Cat#2971, Cell signaling, Danvers, MA, USA), total-mTOR (1:1000, Cat#2972, Cell signaling, Danvers, MA, USA), p-4EBP1 (1:1000, Cat#9455, Cell signaling, Danvers, MA, USA), total-4EBP1 (1:1000, Cat#9452, Cell signaling, Danvers, MA, USA), p-S6 ribosomal protein (1:2000, Cat#5364, Cell signaling, Danvers, MA, USA), S6 ribosomal protein (1:2000, Cat#2317, Cell signaling, Danvers, MA, USA), and β-actin (1:2000, #Cat4967, Cell signaling, Danvers, MA, USA) as a loading control. ..

    Multiple Displacement Amplification:

    Article Title: The PERK-eIF2α-ATF4 Axis Is Involved in Mediating ER-Stress-Induced Ferroptosis via DDIT4-mTORC1 Inhibition and Acetaminophen-Induced Hepatotoxicity.
    Article Snippet: Samples were boiled at 95 ◦C in 2x×Laemmli Sample Buffer (1610737, Bio-Rad, Hercules, CA, USA) for 5 min. Proteins were resolved by SDS-PAGE and transferred to polyvinylidene difluoride membranes (IPVH00010, Millipore, Burlington, MA, USA). .. The membranes were blocked with 5% nonfat milk (BD232100, BD bioscience, San Diego, CA, USA) in phosphate-buffered salineTween 20 (PBS-T), and then the membranes were incubated with primary antibodies as follows: p-PERK (1:1000, Cat#12814, Signalway antibody, Greenbelt, MD, USA), PERK (1:1000, Cat#5683, Cell Signaling, Danvers, MA, USA), p-eIF2α (1:1000, Cat#9721, Cell Signaling, Danvers, MA, USA), eIF2α (1:1000, Cat#9722, Cell Signaling, Danvers, MA, USA), ATF4 (1:1000, Cat#10835-1-AP, Proteintech, Rosemont, IL, USA), GRP78 (1:1000, Cat#ADI-SPA-826, Enzo Life Sciences, Farmingdale, NY, USA), DDIT3 (1:500, Cat#sc7351, Santa Cruz, Dallas, TX, USA), p-IRE1α (1:1000, Cat#NB100-2323, Novus Biologicals, Centennial, CO, USA), total-IRE1α (1:500, Cat#sc-390960, Santa Cruz, Dallas, TX, USA), ATF6 (1:1000, #Cat24169-AP, Proteintech, Rosemont, IL, USA), XBP-1(s) (1:1000, Cat#647502, BioLegend, San Diego, CA, USA), GPX4 (1:2000, Cat#Ab125066, Abcam, Cambridge, UK), SLC7A11 (1:1000, Cat#PA1-16775, Thermo Fisher Scientific, Waltham, MA, USA), MDA (1:1000, Cat#6463, Abcam, Cambridge, UK), 4-HNE (1:1000, Cat#46545, Abcam, Cambridge, UK), CHAC1 (1:1000, Cat#15207-AP, Proteintech, Rosemont, IL, USA), DDIT4 (1:1000, Cat#10638-1-AP, Proteintech, Rosemont, IL, USA), cleaved caspase-3 (1:1000, Cat#9661, Cell signaling, Danvers, MA, USA), caspase-3 (1:1000, Cat#9662, Cell signaling, Danvers, MA, USA), p-mTOR (1:1000, Cat#2971, Cell signaling, Danvers, MA, USA), total-mTOR (1:1000, Cat#2972, Cell signaling, Danvers, MA, USA), p-4EBP1 (1:1000, Cat#9455, Cell signaling, Danvers, MA, USA), total-4EBP1 (1:1000, Cat#9452, Cell signaling, Danvers, MA, USA), p-S6 ribosomal protein (1:2000, Cat#5364, Cell signaling, Danvers, MA, USA), S6 ribosomal protein (1:2000, Cat#2317, Cell signaling, Danvers, MA, USA), and β-actin (1:2000, #Cat4967, Cell signaling, Danvers, MA, USA) as a loading control. ..

    Article Title: The PERK-eIF2α-ATF4 Axis Is Involved in Mediating ER-Stress-Induced Ferroptosis via DDIT4-mTORC1 Inhibition and Acetaminophen-Induced Hepatotoxicity
    Article Snippet: Samples were boiled at 95 °C in 2x×Laemmli Sample Buffer (1610737, Bio-Rad, Hercules, CA, USA) for 5 min. Proteins were resolved by SDS-PAGE and transferred to polyvinylidene difluoride membranes (IPVH00010, Millipore, Burlington, MA, USA). .. The membranes were blocked with 5% nonfat milk (BD232100, BD bioscience, San Diego, CA, USA) in phosphate-buffered saline-Tween 20 (PBS-T), and then the membranes were incubated with primary antibodies as follows: p-PERK (1:1000, Cat#12814, Signalway antibody, Greenbelt, MD, USA), PERK (1:1000, Cat#5683, Cell Signaling, Danvers, MA, USA), p-eIF2α (1:1000, Cat#9721, Cell Signaling, Danvers, MA, USA), eIF2α (1:1000, Cat#9722, Cell Signaling, Danvers, MA, USA), ATF4 (1:1000, Cat#10835-1-AP, Proteintech, Rosemont, IL, USA), GRP78 (1:1000, Cat#ADI-SPA-826, Enzo Life Sciences, Farmingdale, NY, USA), DDIT3 (1:500, Cat#sc-7351, Santa Cruz, Dallas, TX, USA), p-IRE1α (1:1000, Cat#NB100-2323, Novus Biologicals, Centennial, CO, USA), total-IRE1α (1:500, Cat#sc-390960, Santa Cruz, Dallas, TX, USA), ATF6 (1:1000, #Cat24169-AP, Proteintech, Rosemont, IL, USA), XBP-1(s) (1:1000, Cat#647502, BioLegend, San Diego, CA, USA), GPX4 (1:2000, Cat#Ab125066, Abcam, Cambridge, UK), SLC7A11 (1:1000, Cat#PA1-16775, Thermo Fisher Scientific, Waltham, MA, USA), MDA (1:1000, Cat#6463, Abcam, Cambridge, UK), 4-HNE (1:1000, Cat#46545, Abcam, Cambridge, UK), CHAC1 (1:1000, Cat#15207-AP, Proteintech, Rosemont, IL, USA), DDIT4 (1:1000, Cat#10638-1-AP, Proteintech, Rosemont, IL, USA), cleaved caspase-3 (1:1000, Cat#9661, Cell signaling, Danvers, MA, USA), caspase-3 (1:1000, Cat#9662, Cell signaling, Danvers, MA, USA), p-mTOR (1:1000, Cat#2971, Cell signaling, Danvers, MA, USA), total-mTOR (1:1000, Cat#2972, Cell signaling, Danvers, MA, USA), p-4EBP1 (1:1000, Cat#9455, Cell signaling, Danvers, MA, USA), total-4EBP1 (1:1000, Cat#9452, Cell signaling, Danvers, MA, USA), p-S6 ribosomal protein (1:2000, Cat#5364, Cell signaling, Danvers, MA, USA), S6 ribosomal protein (1:2000, Cat#2317, Cell signaling, Danvers, MA, USA), and β-actin (1:2000, #Cat4967, Cell signaling, Danvers, MA, USA) as a loading control. ..

    Control:

    Article Title: The PERK-eIF2α-ATF4 Axis Is Involved in Mediating ER-Stress-Induced Ferroptosis via DDIT4-mTORC1 Inhibition and Acetaminophen-Induced Hepatotoxicity.
    Article Snippet: Samples were boiled at 95 ◦C in 2x×Laemmli Sample Buffer (1610737, Bio-Rad, Hercules, CA, USA) for 5 min. Proteins were resolved by SDS-PAGE and transferred to polyvinylidene difluoride membranes (IPVH00010, Millipore, Burlington, MA, USA). .. The membranes were blocked with 5% nonfat milk (BD232100, BD bioscience, San Diego, CA, USA) in phosphate-buffered salineTween 20 (PBS-T), and then the membranes were incubated with primary antibodies as follows: p-PERK (1:1000, Cat#12814, Signalway antibody, Greenbelt, MD, USA), PERK (1:1000, Cat#5683, Cell Signaling, Danvers, MA, USA), p-eIF2α (1:1000, Cat#9721, Cell Signaling, Danvers, MA, USA), eIF2α (1:1000, Cat#9722, Cell Signaling, Danvers, MA, USA), ATF4 (1:1000, Cat#10835-1-AP, Proteintech, Rosemont, IL, USA), GRP78 (1:1000, Cat#ADI-SPA-826, Enzo Life Sciences, Farmingdale, NY, USA), DDIT3 (1:500, Cat#sc7351, Santa Cruz, Dallas, TX, USA), p-IRE1α (1:1000, Cat#NB100-2323, Novus Biologicals, Centennial, CO, USA), total-IRE1α (1:500, Cat#sc-390960, Santa Cruz, Dallas, TX, USA), ATF6 (1:1000, #Cat24169-AP, Proteintech, Rosemont, IL, USA), XBP-1(s) (1:1000, Cat#647502, BioLegend, San Diego, CA, USA), GPX4 (1:2000, Cat#Ab125066, Abcam, Cambridge, UK), SLC7A11 (1:1000, Cat#PA1-16775, Thermo Fisher Scientific, Waltham, MA, USA), MDA (1:1000, Cat#6463, Abcam, Cambridge, UK), 4-HNE (1:1000, Cat#46545, Abcam, Cambridge, UK), CHAC1 (1:1000, Cat#15207-AP, Proteintech, Rosemont, IL, USA), DDIT4 (1:1000, Cat#10638-1-AP, Proteintech, Rosemont, IL, USA), cleaved caspase-3 (1:1000, Cat#9661, Cell signaling, Danvers, MA, USA), caspase-3 (1:1000, Cat#9662, Cell signaling, Danvers, MA, USA), p-mTOR (1:1000, Cat#2971, Cell signaling, Danvers, MA, USA), total-mTOR (1:1000, Cat#2972, Cell signaling, Danvers, MA, USA), p-4EBP1 (1:1000, Cat#9455, Cell signaling, Danvers, MA, USA), total-4EBP1 (1:1000, Cat#9452, Cell signaling, Danvers, MA, USA), p-S6 ribosomal protein (1:2000, Cat#5364, Cell signaling, Danvers, MA, USA), S6 ribosomal protein (1:2000, Cat#2317, Cell signaling, Danvers, MA, USA), and β-actin (1:2000, #Cat4967, Cell signaling, Danvers, MA, USA) as a loading control. ..

    Article Title: The PERK-eIF2α-ATF4 Axis Is Involved in Mediating ER-Stress-Induced Ferroptosis via DDIT4-mTORC1 Inhibition and Acetaminophen-Induced Hepatotoxicity
    Article Snippet: Samples were boiled at 95 °C in 2x×Laemmli Sample Buffer (1610737, Bio-Rad, Hercules, CA, USA) for 5 min. Proteins were resolved by SDS-PAGE and transferred to polyvinylidene difluoride membranes (IPVH00010, Millipore, Burlington, MA, USA). .. The membranes were blocked with 5% nonfat milk (BD232100, BD bioscience, San Diego, CA, USA) in phosphate-buffered saline-Tween 20 (PBS-T), and then the membranes were incubated with primary antibodies as follows: p-PERK (1:1000, Cat#12814, Signalway antibody, Greenbelt, MD, USA), PERK (1:1000, Cat#5683, Cell Signaling, Danvers, MA, USA), p-eIF2α (1:1000, Cat#9721, Cell Signaling, Danvers, MA, USA), eIF2α (1:1000, Cat#9722, Cell Signaling, Danvers, MA, USA), ATF4 (1:1000, Cat#10835-1-AP, Proteintech, Rosemont, IL, USA), GRP78 (1:1000, Cat#ADI-SPA-826, Enzo Life Sciences, Farmingdale, NY, USA), DDIT3 (1:500, Cat#sc-7351, Santa Cruz, Dallas, TX, USA), p-IRE1α (1:1000, Cat#NB100-2323, Novus Biologicals, Centennial, CO, USA), total-IRE1α (1:500, Cat#sc-390960, Santa Cruz, Dallas, TX, USA), ATF6 (1:1000, #Cat24169-AP, Proteintech, Rosemont, IL, USA), XBP-1(s) (1:1000, Cat#647502, BioLegend, San Diego, CA, USA), GPX4 (1:2000, Cat#Ab125066, Abcam, Cambridge, UK), SLC7A11 (1:1000, Cat#PA1-16775, Thermo Fisher Scientific, Waltham, MA, USA), MDA (1:1000, Cat#6463, Abcam, Cambridge, UK), 4-HNE (1:1000, Cat#46545, Abcam, Cambridge, UK), CHAC1 (1:1000, Cat#15207-AP, Proteintech, Rosemont, IL, USA), DDIT4 (1:1000, Cat#10638-1-AP, Proteintech, Rosemont, IL, USA), cleaved caspase-3 (1:1000, Cat#9661, Cell signaling, Danvers, MA, USA), caspase-3 (1:1000, Cat#9662, Cell signaling, Danvers, MA, USA), p-mTOR (1:1000, Cat#2971, Cell signaling, Danvers, MA, USA), total-mTOR (1:1000, Cat#2972, Cell signaling, Danvers, MA, USA), p-4EBP1 (1:1000, Cat#9455, Cell signaling, Danvers, MA, USA), total-4EBP1 (1:1000, Cat#9452, Cell signaling, Danvers, MA, USA), p-S6 ribosomal protein (1:2000, Cat#5364, Cell signaling, Danvers, MA, USA), S6 ribosomal protein (1:2000, Cat#2317, Cell signaling, Danvers, MA, USA), and β-actin (1:2000, #Cat4967, Cell signaling, Danvers, MA, USA) as a loading control. ..

    Ubiquitin Proteomics:

    Article Title: Acidosis Forces Fatty Acid Uptake and Metabolism in Cancer Cells Regardless of Genotype
    Article Snippet: .. Then, membranes were blocked with 5% skimmed milk diluted in TBS‐0.1% Tween 20 and immunoblotted overnight at 4 °C with specific primary antibodies, P‐p38 (#4511, Cell Signaling, 1:1000), p38 (#9212, Cell Signaling, 1:1000), P‐eIF2α (#9721, Cell Signaling, 1:1000), eIF2α (#2103S, Cell Signaling, 1:1000), P‐IRE1α (NB100‐2323, Novus Biological, 1:1000), IRE1α (#3294, Cell Signaling, 1:1000), Ubiquitin (#3936, Cell Signaling, 1:1000) and β‐actin (A5441, Merck, 1:10 000). .. After several washes with TTBS, membranes were incubated for 1 h at RT with horseradish peroxidase (HRP)‐conjugated secondary antibodies (Jackson Immunoresearch), and chemiluminescent signals were revealed by using ECL Western Blotting Detection Kit (GE Healthcare) on X‐ray films in a dark chamber.

    Article Title: Acidosis Forces Fatty Acid Uptake and Metabolism in Cancer Cells Regardless of Genotype.
    Article Snippet: .. Then, membranes were blocked with 5% skimmed milk diluted in TBS0.1% Tween 20 and immunoblotted overnight at 4 °C with specific primary antibodies, P-p38 (#4511, Cell Signaling, 1:1000), p38 (#9212, Cell Signaling, 1:1000), P-eIF2α (#9721, Cell Signaling, 1:1000), eIF2α (#2103S, Cell Signaling, 1:1000), P-IRE1α (NB100-2323, Novus Biological, 1:1000), IRE1α (#3294, Cell Signaling, 1:1000), Ubiquitin (#3936, Cell Signaling, 1:1000) and β-actin (A5441, Merck, 1:10 000). .. After several washes with TTBS, membranes were incubated for 1 h at RT with horseradish peroxidase (HRP)-conjugated secondary antibodies (Jackson Immunoresearch), and chemiluminescent signals were revealed by using ECL Western Blotting Detection Kit (GE Healthcare) on X-ray films in a dark chamber.

    Western Blot:

    Article Title: Licorice attenuates cisplatin-induced hepatotoxicity by alleviating endoplasmic reticulum stress and apoptosis
    Article Snippet: .. WB analysis was performed as previously described ( ) using specific primary antibodies against CHOP (1:1000, 15204-1-AP, Proteintech), ATF4 (1:1000, BM5179, Boster), ATF6 (1:1000, A00655, Boster), GRP78 (1:1000, BA 2042, Boster), p-IRE1α (1:1000, NB100-2323, Novus), p-eIF2α (1:1000, BM3942, Boster), Bcl-2 (1:1000, A00040-1, Boster), Bax (1:1000, A00183, Boster), cleaved caspase-3 (1:1000, #9664T, Cell Signaling Technology), caspase-12 (1:1000, BA3142, Boster), cleaved caspase-8 (1:5000, ab108333, Abcam), p-PERK (1:1000, abs137056, Absin) and β-actin (1:5000, 20536-1-AP, Proteintech). .. Using HRP-conjugated Rabbit Anti-Goat IgG (H + L) (1:1000, SA0000-1-4, Proteintech) as the secondary antibody.

    Article Title: Treatment and prevention of pulmonary injury
    Article Snippet: ER stress was analyzed by measuring Grp78 using a Grp78/Bip ELISA kit (ADI-900-214, Enzo Life Sciences) according to the manufacturer's instructions. .. Immunoblot was performed using primary antibodies at 1:1,000 dilution against CHOP (2895S), ATF4 (11815S), total IRE1α (3294S), p-PERK (3179S), p-eIF2α (9721S), total eIF2α (9722S) (all Cell Signalling Technology); ATF6 (sc22799), total PERK (sc13073), Grp78 (sc13968) (all Santa Cruz Biotechnology); and p-IRE1α (NB100-2323, Novus Biologicals) as described previously [8]. .. Bands were detected by a chemiluminescent HRP substrate (Millipore).

    Saline:

    Article Title: The PERK-eIF2α-ATF4 Axis Is Involved in Mediating ER-Stress-Induced Ferroptosis via DDIT4-mTORC1 Inhibition and Acetaminophen-Induced Hepatotoxicity
    Article Snippet: Samples were boiled at 95 °C in 2x×Laemmli Sample Buffer (1610737, Bio-Rad, Hercules, CA, USA) for 5 min. Proteins were resolved by SDS-PAGE and transferred to polyvinylidene difluoride membranes (IPVH00010, Millipore, Burlington, MA, USA). .. The membranes were blocked with 5% nonfat milk (BD232100, BD bioscience, San Diego, CA, USA) in phosphate-buffered saline-Tween 20 (PBS-T), and then the membranes were incubated with primary antibodies as follows: p-PERK (1:1000, Cat#12814, Signalway antibody, Greenbelt, MD, USA), PERK (1:1000, Cat#5683, Cell Signaling, Danvers, MA, USA), p-eIF2α (1:1000, Cat#9721, Cell Signaling, Danvers, MA, USA), eIF2α (1:1000, Cat#9722, Cell Signaling, Danvers, MA, USA), ATF4 (1:1000, Cat#10835-1-AP, Proteintech, Rosemont, IL, USA), GRP78 (1:1000, Cat#ADI-SPA-826, Enzo Life Sciences, Farmingdale, NY, USA), DDIT3 (1:500, Cat#sc-7351, Santa Cruz, Dallas, TX, USA), p-IRE1α (1:1000, Cat#NB100-2323, Novus Biologicals, Centennial, CO, USA), total-IRE1α (1:500, Cat#sc-390960, Santa Cruz, Dallas, TX, USA), ATF6 (1:1000, #Cat24169-AP, Proteintech, Rosemont, IL, USA), XBP-1(s) (1:1000, Cat#647502, BioLegend, San Diego, CA, USA), GPX4 (1:2000, Cat#Ab125066, Abcam, Cambridge, UK), SLC7A11 (1:1000, Cat#PA1-16775, Thermo Fisher Scientific, Waltham, MA, USA), MDA (1:1000, Cat#6463, Abcam, Cambridge, UK), 4-HNE (1:1000, Cat#46545, Abcam, Cambridge, UK), CHAC1 (1:1000, Cat#15207-AP, Proteintech, Rosemont, IL, USA), DDIT4 (1:1000, Cat#10638-1-AP, Proteintech, Rosemont, IL, USA), cleaved caspase-3 (1:1000, Cat#9661, Cell signaling, Danvers, MA, USA), caspase-3 (1:1000, Cat#9662, Cell signaling, Danvers, MA, USA), p-mTOR (1:1000, Cat#2971, Cell signaling, Danvers, MA, USA), total-mTOR (1:1000, Cat#2972, Cell signaling, Danvers, MA, USA), p-4EBP1 (1:1000, Cat#9455, Cell signaling, Danvers, MA, USA), total-4EBP1 (1:1000, Cat#9452, Cell signaling, Danvers, MA, USA), p-S6 ribosomal protein (1:2000, Cat#5364, Cell signaling, Danvers, MA, USA), S6 ribosomal protein (1:2000, Cat#2317, Cell signaling, Danvers, MA, USA), and β-actin (1:2000, #Cat4967, Cell signaling, Danvers, MA, USA) as a loading control. ..

    other:

    Article Title: Lipid Metabolic Changes and Mitochondrial Stress in Ethanol-Treated Alveolar Type II Epithelial Cells: Initial Events Leading to Alcoholic Chronic Lung Disease
    Article Snippet: Antibodies against inositol-requiring transmembrane kinase/endoribonuclease 1α (IRE1α; 110 kDa; Cat # NB100-2324), and p-IRE1α (Ser 724) (110 kDa; Cat # NB100-232) were obtained from Novus Biologicals (Littleton, CO, USA).



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    A . Boxplot for data standardization evaluation. B . Principal component analysis scatter plot. C . Volcano plot of differentially expressed genes. D . Heatmap of differentially expressed genes. Ea. GO cellular component (CC) enrichment bubble plot. Eb. GO molecular function (MF) enrichment bubble plot. Ec. GO biological process (BP) enrichment bubble plot. Fa. KEGG pathway enrichment bubble plot. Fb. KEGG pathway enrichment lollipop plot. G . Correlation pie chart of key regulatory factors <t>(IRE1α,</t> XBP-1, p38, SP1, ZEB1, PKP3, Rb, E2F1, Cyclin, SHP2, BRD4). H . Scatter plot of E2F1-CDK1 co-regulation. I. Scatter plot of NLRP3-GSDMD pyroptosis axis.
    P Ire1α, supplied by Wuhan Sanying Biotechnology, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    phosphorylated ire1α at ser724 p ire1α  (Novus Biologicals)
    96
    Novus Biologicals phosphorylated ire1α at ser724 p ire1α
    A . Boxplot for data standardization evaluation. B . Principal component analysis scatter plot. C . Volcano plot of differentially expressed genes. D . Heatmap of differentially expressed genes. Ea. GO cellular component (CC) enrichment bubble plot. Eb. GO molecular function (MF) enrichment bubble plot. Ec. GO biological process (BP) enrichment bubble plot. Fa. KEGG pathway enrichment bubble plot. Fb. KEGG pathway enrichment lollipop plot. G . Correlation pie chart of key regulatory factors <t>(IRE1α,</t> XBP-1, p38, SP1, ZEB1, PKP3, Rb, E2F1, Cyclin, SHP2, BRD4). H . Scatter plot of E2F1-CDK1 co-regulation. I. Scatter plot of NLRP3-GSDMD pyroptosis axis.
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    anti ire1α  (Novus Biologicals)
    96
    Novus Biologicals anti ire1α
    A . Boxplot for data standardization evaluation. B . Principal component analysis scatter plot. C . Volcano plot of differentially expressed genes. D . Heatmap of differentially expressed genes. Ea. GO cellular component (CC) enrichment bubble plot. Eb. GO molecular function (MF) enrichment bubble plot. Ec. GO biological process (BP) enrichment bubble plot. Fa. KEGG pathway enrichment bubble plot. Fb. KEGG pathway enrichment lollipop plot. G . Correlation pie chart of key regulatory factors <t>(IRE1α,</t> XBP-1, p38, SP1, ZEB1, PKP3, Rb, E2F1, Cyclin, SHP2, BRD4). H . Scatter plot of E2F1-CDK1 co-regulation. I. Scatter plot of NLRP3-GSDMD pyroptosis axis.
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    p ire1α  (Wanleibio)
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    Wanleibio p ire1α
    A . Boxplot for data standardization evaluation. B . Principal component analysis scatter plot. C . Volcano plot of differentially expressed genes. D . Heatmap of differentially expressed genes. Ea. GO cellular component (CC) enrichment bubble plot. Eb. GO molecular function (MF) enrichment bubble plot. Ec. GO biological process (BP) enrichment bubble plot. Fa. KEGG pathway enrichment bubble plot. Fb. KEGG pathway enrichment lollipop plot. G . Correlation pie chart of key regulatory factors <t>(IRE1α,</t> XBP-1, p38, SP1, ZEB1, PKP3, Rb, E2F1, Cyclin, SHP2, BRD4). H . Scatter plot of E2F1-CDK1 co-regulation. I. Scatter plot of NLRP3-GSDMD pyroptosis axis.
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    p ire1α  (Bioss)
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    Bioss p ire1α
    NHE7 activates ER stress pathways in vitro and in vivo. A WB analysis of ER stress markers (p-PERK, <t>p-IRE1α,</t> ATF6) in Ishikawa, HEC-1-A, and HEC-1-B cells following NHE7 overexpressing. B WB analysis of ER stress markers (p-PERK, p-IRE1α, ATF6) in Ishikawa, HEC-1-A, and HEC-1-B cells following NHE7 knockdown. C IHC staining of p-IRE1α and ATF6 in xenograft tumor tissues with NHE7 overexpression. Scale bars, 50 μm (400 ×) and 100 μm (200 ×). D Correlation analysis between NHE7 and ER stress markers (p-IRE1α, ATF6) in clinical EC samples by IHC. Scale bars, 50 μm (400 ×) and 100 μm (200 ×). At least three independent experiments were performed on the assays. Error bars represent the mean ± SD ( n = 3). * P < 0.05,** P < 0.01
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    p ire1α  (Santa Cruz Biotechnology)
    96
    Santa Cruz Biotechnology p ire1α
    The mRNA levels of GRP78/BiP in LoVo ( A ) and HCT116 ( B ) cells after ART knockdown. * P < 0.01 by 1-way ANOVA with Tukeys HSD test (mean ± SEM, n = 3). ( C ) The protein expression levels of GRP78/BiP in pHBLV, KRAS-WT, G12D, and G13D cells treated with ART1 inhibitor MIBG for 12, 24, 36, and 48 hours. * P < 0.01 by t test (mean ± SEM, n = 3). Effects of GRP78/BiP arginine mono-ADP-ribosylation modification by MIBG on the binding of GRP78/BiP to its receptors PERK ( D ), <t>IRE1α</t> ( E ), and ATF6 ( F ) in KRAS-WT, G12D, and G13D cell lines. * P < 0.01 by t test (mean ± SEM, n = 3). ( G and H ) The expression levels of key proteins in the UPR signaling pathways IRE1α/XBP1/TFAF2/JNK, PERK/eIF2α/ATF4, and ATF6/S1P/S2P/CHOP in pHBLV, KRAS-WT, G12D, and G13D cells treated with MIBG. * P < 0.01 by t test (mean ± SEM, n = 3). ( I ) The effect of ART1 inhibitor MIBG on the expression of cleaved caspase-3 in KRAS-WT, G12D, and G13D cells. * P < 0.01 by t test (mean ± SEM, n = 3).
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    Image Search Results


    A . Boxplot for data standardization evaluation. B . Principal component analysis scatter plot. C . Volcano plot of differentially expressed genes. D . Heatmap of differentially expressed genes. Ea. GO cellular component (CC) enrichment bubble plot. Eb. GO molecular function (MF) enrichment bubble plot. Ec. GO biological process (BP) enrichment bubble plot. Fa. KEGG pathway enrichment bubble plot. Fb. KEGG pathway enrichment lollipop plot. G . Correlation pie chart of key regulatory factors (IRE1α, XBP-1, p38, SP1, ZEB1, PKP3, Rb, E2F1, Cyclin, SHP2, BRD4). H . Scatter plot of E2F1-CDK1 co-regulation. I. Scatter plot of NLRP3-GSDMD pyroptosis axis.

    Journal: Scientific Reports

    Article Title: SHP2 improves ovarian morphology and steroidogenic function in a rat PCOS model by modulating IRE1α/XBP1/NLRP3-mediated granulosa cell pyroptosis

    doi: 10.1038/s41598-026-43536-2

    Figure Lengend Snippet: A . Boxplot for data standardization evaluation. B . Principal component analysis scatter plot. C . Volcano plot of differentially expressed genes. D . Heatmap of differentially expressed genes. Ea. GO cellular component (CC) enrichment bubble plot. Eb. GO molecular function (MF) enrichment bubble plot. Ec. GO biological process (BP) enrichment bubble plot. Fa. KEGG pathway enrichment bubble plot. Fb. KEGG pathway enrichment lollipop plot. G . Correlation pie chart of key regulatory factors (IRE1α, XBP-1, p38, SP1, ZEB1, PKP3, Rb, E2F1, Cyclin, SHP2, BRD4). H . Scatter plot of E2F1-CDK1 co-regulation. I. Scatter plot of NLRP3-GSDMD pyroptosis axis.

    Article Snippet: Antibodies against p-SHP2, p-IRE1α, XBP-1s, p-SP1, ZEB1, PKP3, nuclear E2F1, CyclinB1, NLRP3, GSDMD, and GAPDH were purchased from Wuhan Sanying Biotechnology Co., Ltd. IgG secondary antibodies and ECL hypersensitive luminescent solution were purchased from Beijing Biosynthesis Biotechnology Co., Ltd.

    Techniques:

    SHP2 Alleviates PCOS Phenotypes via IRE1α/XBP1/NLRP3 and ZEB1/PKP3-Mediated Regulation. (A) Western blot detection of SHP2-regulated IRE1α/XBP1/ZEB1/PKP3 protein pathways. (B) Quantitative analysis of protein expression levels.* p < 0.05, ** p < 0.01, ns indicates P > 0.05; data are represented as the mean ± SD.

    Journal: Scientific Reports

    Article Title: SHP2 improves ovarian morphology and steroidogenic function in a rat PCOS model by modulating IRE1α/XBP1/NLRP3-mediated granulosa cell pyroptosis

    doi: 10.1038/s41598-026-43536-2

    Figure Lengend Snippet: SHP2 Alleviates PCOS Phenotypes via IRE1α/XBP1/NLRP3 and ZEB1/PKP3-Mediated Regulation. (A) Western blot detection of SHP2-regulated IRE1α/XBP1/ZEB1/PKP3 protein pathways. (B) Quantitative analysis of protein expression levels.* p < 0.05, ** p < 0.01, ns indicates P > 0.05; data are represented as the mean ± SD.

    Article Snippet: Antibodies against p-SHP2, p-IRE1α, XBP-1s, p-SP1, ZEB1, PKP3, nuclear E2F1, CyclinB1, NLRP3, GSDMD, and GAPDH were purchased from Wuhan Sanying Biotechnology Co., Ltd. IgG secondary antibodies and ECL hypersensitive luminescent solution were purchased from Beijing Biosynthesis Biotechnology Co., Ltd.

    Techniques: Western Blot, Expressing

    SHP2 improves ovarian morphology and steroidogenic function in a rat PCOS model by regulating the IRE1α/XBP1/NLRP3 and ZEB1/PKP3 signaling pathways, thereby influencing granulosa cell pyroptosis and proliferation.

    Journal: Scientific Reports

    Article Title: SHP2 improves ovarian morphology and steroidogenic function in a rat PCOS model by modulating IRE1α/XBP1/NLRP3-mediated granulosa cell pyroptosis

    doi: 10.1038/s41598-026-43536-2

    Figure Lengend Snippet: SHP2 improves ovarian morphology and steroidogenic function in a rat PCOS model by regulating the IRE1α/XBP1/NLRP3 and ZEB1/PKP3 signaling pathways, thereby influencing granulosa cell pyroptosis and proliferation.

    Article Snippet: Antibodies against p-SHP2, p-IRE1α, XBP-1s, p-SP1, ZEB1, PKP3, nuclear E2F1, CyclinB1, NLRP3, GSDMD, and GAPDH were purchased from Wuhan Sanying Biotechnology Co., Ltd. IgG secondary antibodies and ECL hypersensitive luminescent solution were purchased from Beijing Biosynthesis Biotechnology Co., Ltd.

    Techniques: Protein-Protein interactions

    NHE7 activates ER stress pathways in vitro and in vivo. A WB analysis of ER stress markers (p-PERK, p-IRE1α, ATF6) in Ishikawa, HEC-1-A, and HEC-1-B cells following NHE7 overexpressing. B WB analysis of ER stress markers (p-PERK, p-IRE1α, ATF6) in Ishikawa, HEC-1-A, and HEC-1-B cells following NHE7 knockdown. C IHC staining of p-IRE1α and ATF6 in xenograft tumor tissues with NHE7 overexpression. Scale bars, 50 μm (400 ×) and 100 μm (200 ×). D Correlation analysis between NHE7 and ER stress markers (p-IRE1α, ATF6) in clinical EC samples by IHC. Scale bars, 50 μm (400 ×) and 100 μm (200 ×). At least three independent experiments were performed on the assays. Error bars represent the mean ± SD ( n = 3). * P < 0.05,** P < 0.01

    Journal: Apoptosis

    Article Title: Hypoxic glycolysis-driven histone lactylation activates NHE7 to promote endometrial cancer progression via COX6C-mediated endoplasmic reticulum stress

    doi: 10.1007/s10495-026-02262-w

    Figure Lengend Snippet: NHE7 activates ER stress pathways in vitro and in vivo. A WB analysis of ER stress markers (p-PERK, p-IRE1α, ATF6) in Ishikawa, HEC-1-A, and HEC-1-B cells following NHE7 overexpressing. B WB analysis of ER stress markers (p-PERK, p-IRE1α, ATF6) in Ishikawa, HEC-1-A, and HEC-1-B cells following NHE7 knockdown. C IHC staining of p-IRE1α and ATF6 in xenograft tumor tissues with NHE7 overexpression. Scale bars, 50 μm (400 ×) and 100 μm (200 ×). D Correlation analysis between NHE7 and ER stress markers (p-IRE1α, ATF6) in clinical EC samples by IHC. Scale bars, 50 μm (400 ×) and 100 μm (200 ×). At least three independent experiments were performed on the assays. Error bars represent the mean ± SD ( n = 3). * P < 0.05,** P < 0.01

    Article Snippet: p-IRE1α , BIOSS , bs-16698R , WB/IHC.

    Techniques: In Vitro, In Vivo, Knockdown, Immunohistochemistry, Over Expression

    NHE7 enhances malignant phenotypes and stemness in EC cells by activating the ER stress pathway. A MTT assays were performed to evaluate the effect of 4-PBA concentration on cell viability in Ishikawa cells. Ishikawa cells were divided into pCDH + Control, NHE7 + control and NHE7 + 4-PBA (ER stress inhibitor). B The expressions of p-IRE1α, IRE1α and ATF6 were examined in Ishikawa cells through western blot assay. C The proliferation ability of Ishikawa cells was detected using the MTT assay. D The apoptosis levels of Ishikawa cells were detected using flow cytometry. E The expressions of apoptosis-related markers (cleaved PARP and cleaved Caspase-3) in Ishikawa cells were detected by WB assay. F The colony-forming capacity of Ishikawa cells was tested using a colony formation assay. G The migration and invasion abilities of Ishikawa cells were detected through transwell experiments. Scale bars, 100 μm (100 ×). H Tumor sphere-forming assay showed the effects of NHE7 overexpression and 4-PBA on the tumor sphere formation of Ishikawa cells. I WB analysis assessed the expression levels of EMT markers (E-cadherin, N-cadherin, and Vimentin) and stemness-associated proteins (OCT4, Nanog, and SOX2). At least three independent experiments were performed on the assays. Error bars represent the mean ± SD ( n = 3). * P < 0.05, ** P < 0.01

    Journal: Apoptosis

    Article Title: Hypoxic glycolysis-driven histone lactylation activates NHE7 to promote endometrial cancer progression via COX6C-mediated endoplasmic reticulum stress

    doi: 10.1007/s10495-026-02262-w

    Figure Lengend Snippet: NHE7 enhances malignant phenotypes and stemness in EC cells by activating the ER stress pathway. A MTT assays were performed to evaluate the effect of 4-PBA concentration on cell viability in Ishikawa cells. Ishikawa cells were divided into pCDH + Control, NHE7 + control and NHE7 + 4-PBA (ER stress inhibitor). B The expressions of p-IRE1α, IRE1α and ATF6 were examined in Ishikawa cells through western blot assay. C The proliferation ability of Ishikawa cells was detected using the MTT assay. D The apoptosis levels of Ishikawa cells were detected using flow cytometry. E The expressions of apoptosis-related markers (cleaved PARP and cleaved Caspase-3) in Ishikawa cells were detected by WB assay. F The colony-forming capacity of Ishikawa cells was tested using a colony formation assay. G The migration and invasion abilities of Ishikawa cells were detected through transwell experiments. Scale bars, 100 μm (100 ×). H Tumor sphere-forming assay showed the effects of NHE7 overexpression and 4-PBA on the tumor sphere formation of Ishikawa cells. I WB analysis assessed the expression levels of EMT markers (E-cadherin, N-cadherin, and Vimentin) and stemness-associated proteins (OCT4, Nanog, and SOX2). At least three independent experiments were performed on the assays. Error bars represent the mean ± SD ( n = 3). * P < 0.05, ** P < 0.01

    Article Snippet: p-IRE1α , BIOSS , bs-16698R , WB/IHC.

    Techniques: Concentration Assay, Control, Western Blot, MTT Assay, Flow Cytometry, Colony Assay, Migration, Over Expression, Expressing

    NHE7 enhances OXPHOS-induced ER stress by upregulating COX6C expression in EC cells. A WB was used to detect the transfection efficiency of COX6C knockdown in Ishikawa cells. B WB was used to analyze the expression of NHE7 and COX6C in Ishikawa cells after COX6C silencing combined with NHE7 overexpression. C ELISA was used to determine the level of ATP. D Intracellular ROS levels were detected by flow cytometry. E The expression levels of p-IRE1α, IRE1α, and ATF6 were examined in Ishikawa cells. F The apoptosis levels of Ishikawa cells were detected using flow cytometry. G The expressions of apoptosis-related markers (cleaved PARP and cleaved Caspase-3) in Ishikawa cells were detected by WB assay. H The colony-forming capacity of Ishikawa cells was tested by colony formation assay. I The migration and invasion abilities of Ishikawa cells were detected through transwell experiments. Scale bar, 100 μm (100 ×). J Tumor sphere-forming assay showed the effects on the tumor sphere formation of Ishikawa cells. At least three independent experiments were performed on the assays. Error bars represent the mean ± SD ( n = 3). * P < 0.05,** P < 0.01

    Journal: Apoptosis

    Article Title: Hypoxic glycolysis-driven histone lactylation activates NHE7 to promote endometrial cancer progression via COX6C-mediated endoplasmic reticulum stress

    doi: 10.1007/s10495-026-02262-w

    Figure Lengend Snippet: NHE7 enhances OXPHOS-induced ER stress by upregulating COX6C expression in EC cells. A WB was used to detect the transfection efficiency of COX6C knockdown in Ishikawa cells. B WB was used to analyze the expression of NHE7 and COX6C in Ishikawa cells after COX6C silencing combined with NHE7 overexpression. C ELISA was used to determine the level of ATP. D Intracellular ROS levels were detected by flow cytometry. E The expression levels of p-IRE1α, IRE1α, and ATF6 were examined in Ishikawa cells. F The apoptosis levels of Ishikawa cells were detected using flow cytometry. G The expressions of apoptosis-related markers (cleaved PARP and cleaved Caspase-3) in Ishikawa cells were detected by WB assay. H The colony-forming capacity of Ishikawa cells was tested by colony formation assay. I The migration and invasion abilities of Ishikawa cells were detected through transwell experiments. Scale bar, 100 μm (100 ×). J Tumor sphere-forming assay showed the effects on the tumor sphere formation of Ishikawa cells. At least three independent experiments were performed on the assays. Error bars represent the mean ± SD ( n = 3). * P < 0.05,** P < 0.01

    Article Snippet: p-IRE1α , BIOSS , bs-16698R , WB/IHC.

    Techniques: Expressing, Transfection, Knockdown, Over Expression, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Colony Assay, Migration

    COX6C triggers ER stress and enhances the malignant phenotypes of EC cells by upregulating ROS levels. A Intracellular ROS levels were detected by flow cytometry in Ishikawa cells with COX6C overexpression combined with the antioxidant N-acetylcysteine (NAC). B The expression levels of p-IRE1α, IRE1α, and ATF6 were examined in Ishikawa cells. C The proliferation ability of Ishikawa cells was detected using the MTT assay. D The apoptosis levels of Ishikawa cells were detected using flow cytometry. E The expressions of apoptosis-related markers (cleaved PARP and cleaved Caspase-3) in Ishikawa cells were detected by WB assay. F The colony-forming capacity of Ishikawa cells was tested by colony formation assay. G The migration and invasion abilities of Ishikawa cells were detected through transwell experiments. Scale bar, 100 μm (100 ×). At least three independent experiments were performed on the assays. Error bars represent the mean ± SD ( n = 3). * P < 0.05,** P < 0.01

    Journal: Apoptosis

    Article Title: Hypoxic glycolysis-driven histone lactylation activates NHE7 to promote endometrial cancer progression via COX6C-mediated endoplasmic reticulum stress

    doi: 10.1007/s10495-026-02262-w

    Figure Lengend Snippet: COX6C triggers ER stress and enhances the malignant phenotypes of EC cells by upregulating ROS levels. A Intracellular ROS levels were detected by flow cytometry in Ishikawa cells with COX6C overexpression combined with the antioxidant N-acetylcysteine (NAC). B The expression levels of p-IRE1α, IRE1α, and ATF6 were examined in Ishikawa cells. C The proliferation ability of Ishikawa cells was detected using the MTT assay. D The apoptosis levels of Ishikawa cells were detected using flow cytometry. E The expressions of apoptosis-related markers (cleaved PARP and cleaved Caspase-3) in Ishikawa cells were detected by WB assay. F The colony-forming capacity of Ishikawa cells was tested by colony formation assay. G The migration and invasion abilities of Ishikawa cells were detected through transwell experiments. Scale bar, 100 μm (100 ×). At least three independent experiments were performed on the assays. Error bars represent the mean ± SD ( n = 3). * P < 0.05,** P < 0.01

    Article Snippet: p-IRE1α , BIOSS , bs-16698R , WB/IHC.

    Techniques: Flow Cytometry, Over Expression, Expressing, MTT Assay, Colony Assay, Migration

    NHE7 facilitates EC tumor growth by upregulating COX6C expression in vivo. A WB analysis was used to detect the protein expression levels of NHE7 and COX6C in Ishikawa cells overexpressing NHE7, and in Ishikawa cells with concurrent NHE7 overexpression and COX6C knockdown. B Stable Ishikawa cell lines were established for the following conditions: NHE7 overexpression alone, and concurrent NHE7 overexpression with COX6C knockdown, along with their respective control cells. A xenograft tumor model was then generated using these cells. C Tumor growth was monitored and recorded, and D tumor weight was measured at the endpoint. E WB assay was performed to assess the effects of NHE7 overexpression alone or in combination with COX6C knockdown on the levels of apoptosis markers c-PARP and C-Caspase3 in xenograft tumor tissues. F HE staining was performed to assess histopathological damage in xenograft tumor tissues following NHE7 overexpression, either alone or in combination with COX6C knockdown. IHC was performed to evaluate the impact of NHE7 overexpression, either alone or in combination with COX6C knockdown, on the expression of Ki67, E-cadherin, N-cadherin, Vimentin, OCT4, Nanog, SOX2, p-IRE1α, IRE1α, and ATF6 in xenograft tumor tissues. Scale bar, 50 μm (400 ×) and 100 μm (200 ×). At least three independent experiments were performed on the assays. Error bars represent the mean ± SD ( n = 3). * P < 0.05,** P < 0.01

    Journal: Apoptosis

    Article Title: Hypoxic glycolysis-driven histone lactylation activates NHE7 to promote endometrial cancer progression via COX6C-mediated endoplasmic reticulum stress

    doi: 10.1007/s10495-026-02262-w

    Figure Lengend Snippet: NHE7 facilitates EC tumor growth by upregulating COX6C expression in vivo. A WB analysis was used to detect the protein expression levels of NHE7 and COX6C in Ishikawa cells overexpressing NHE7, and in Ishikawa cells with concurrent NHE7 overexpression and COX6C knockdown. B Stable Ishikawa cell lines were established for the following conditions: NHE7 overexpression alone, and concurrent NHE7 overexpression with COX6C knockdown, along with their respective control cells. A xenograft tumor model was then generated using these cells. C Tumor growth was monitored and recorded, and D tumor weight was measured at the endpoint. E WB assay was performed to assess the effects of NHE7 overexpression alone or in combination with COX6C knockdown on the levels of apoptosis markers c-PARP and C-Caspase3 in xenograft tumor tissues. F HE staining was performed to assess histopathological damage in xenograft tumor tissues following NHE7 overexpression, either alone or in combination with COX6C knockdown. IHC was performed to evaluate the impact of NHE7 overexpression, either alone or in combination with COX6C knockdown, on the expression of Ki67, E-cadherin, N-cadherin, Vimentin, OCT4, Nanog, SOX2, p-IRE1α, IRE1α, and ATF6 in xenograft tumor tissues. Scale bar, 50 μm (400 ×) and 100 μm (200 ×). At least three independent experiments were performed on the assays. Error bars represent the mean ± SD ( n = 3). * P < 0.05,** P < 0.01

    Article Snippet: p-IRE1α , BIOSS , bs-16698R , WB/IHC.

    Techniques: Expressing, In Vivo, Over Expression, Knockdown, Control, Generated, Staining

    The mRNA levels of GRP78/BiP in LoVo ( A ) and HCT116 ( B ) cells after ART knockdown. * P < 0.01 by 1-way ANOVA with Tukeys HSD test (mean ± SEM, n = 3). ( C ) The protein expression levels of GRP78/BiP in pHBLV, KRAS-WT, G12D, and G13D cells treated with ART1 inhibitor MIBG for 12, 24, 36, and 48 hours. * P < 0.01 by t test (mean ± SEM, n = 3). Effects of GRP78/BiP arginine mono-ADP-ribosylation modification by MIBG on the binding of GRP78/BiP to its receptors PERK ( D ), IRE1α ( E ), and ATF6 ( F ) in KRAS-WT, G12D, and G13D cell lines. * P < 0.01 by t test (mean ± SEM, n = 3). ( G and H ) The expression levels of key proteins in the UPR signaling pathways IRE1α/XBP1/TFAF2/JNK, PERK/eIF2α/ATF4, and ATF6/S1P/S2P/CHOP in pHBLV, KRAS-WT, G12D, and G13D cells treated with MIBG. * P < 0.01 by t test (mean ± SEM, n = 3). ( I ) The effect of ART1 inhibitor MIBG on the expression of cleaved caspase-3 in KRAS-WT, G12D, and G13D cells. * P < 0.01 by t test (mean ± SEM, n = 3).

    Journal: JCI Insight

    Article Title: The critical role of GRP78/BiP MARylation in ER stress of KRAS-mutant colorectal cancer

    doi: 10.1172/jci.insight.182809

    Figure Lengend Snippet: The mRNA levels of GRP78/BiP in LoVo ( A ) and HCT116 ( B ) cells after ART knockdown. * P < 0.01 by 1-way ANOVA with Tukeys HSD test (mean ± SEM, n = 3). ( C ) The protein expression levels of GRP78/BiP in pHBLV, KRAS-WT, G12D, and G13D cells treated with ART1 inhibitor MIBG for 12, 24, 36, and 48 hours. * P < 0.01 by t test (mean ± SEM, n = 3). Effects of GRP78/BiP arginine mono-ADP-ribosylation modification by MIBG on the binding of GRP78/BiP to its receptors PERK ( D ), IRE1α ( E ), and ATF6 ( F ) in KRAS-WT, G12D, and G13D cell lines. * P < 0.01 by t test (mean ± SEM, n = 3). ( G and H ) The expression levels of key proteins in the UPR signaling pathways IRE1α/XBP1/TFAF2/JNK, PERK/eIF2α/ATF4, and ATF6/S1P/S2P/CHOP in pHBLV, KRAS-WT, G12D, and G13D cells treated with MIBG. * P < 0.01 by t test (mean ± SEM, n = 3). ( I ) The effect of ART1 inhibitor MIBG on the expression of cleaved caspase-3 in KRAS-WT, G12D, and G13D cells. * P < 0.01 by t test (mean ± SEM, n = 3).

    Article Snippet: The following antibodies were used: ATF6 (catalog 3192), PERK (catalog 65880), IRE1α (catalog 3294), eIF2α (catalog 5324), p-eIF2α (catalog 3398) (all Cell Signaling Technology); GRP78/BiP (catalog sc-13539), HSC70 (catalog sc-24), Bcl2 (catalog sc-7382) (all Santa Cruz Biotechnology); p-IRE1α (catalog ab124945), XBP1u/s (catalog ab220783) (both Abcam); p-PERK (catalog DF7576, Affinity Biosciences); CHOP (catalog NB600-1335SS, Novus Biotechnology); ART1 (catalog A10103, Abclonal Technology Co., Ltd.); Caspase-3 (catalog WL02117), PARP/cleaved-PARP (catalog WL01932) (both Wanlei Biotechnology); β-actin (catalog 6609-1-Ig), Ki67 (catalog 27309-1-AP) (both Proteintech).

    Techniques: Knockdown, Expressing, Modification, Binding Assay, Protein-Protein interactions

    ( A ) The position of arginines 470, 492, and 214 on the GRP78 protein. ( B ) Three-dimensional protein structure of GRP78 after arginines 470, 492, and 214 were mutated to lysine. ( C ) Effects of downstream UPR signaling pathways after GRP78/BiP R470+492K and GRP78/BiP R214K plasmids transfected into G12D cells. * P < 0.01 by 1-way ANOVA with Tukey’s HSD test (mean ± SEM, n = 3). ( D ) Schematic diagram of arginine–mono-ADP-ribosylated GRP78/BiP involved in the regulation of ER homeostasis under the state of ER stress caused by KRAS mutation: ART1 catalyzes the arginine mono-ADP-ribosylation of GRP78/BiP to inactivate it and release the receptors bound to it. The downstream UPR signaling pathway is continuously activated to help maintain the homeostasis of the ER and the survival of tumor cells. ( E ) Schematic diagram of the effect of interference with arginine–mono-ADP-ribosylated GRP78/BiP on the UPR signaling pathway. Interfering with GRP78/BiP arginine mono-ADP-ribosylation modification can downregulate the expression of GRP78/BiP; limited GRP78/BiP can bind to unfolded protein and sensors PERK and IRE1α, inhibiting the IRE1α/XBP1/TFAF2/JNK pathway. The PERK/eIF2α/ATF4 signaling pathway further leads to the reduction of GRP78 expression, damages the ER stress regulation mechanism, increases the pressure of the internal environment, hinders the growth of tumor cells, and the cells tend to undergo apoptosis.

    Journal: JCI Insight

    Article Title: The critical role of GRP78/BiP MARylation in ER stress of KRAS-mutant colorectal cancer

    doi: 10.1172/jci.insight.182809

    Figure Lengend Snippet: ( A ) The position of arginines 470, 492, and 214 on the GRP78 protein. ( B ) Three-dimensional protein structure of GRP78 after arginines 470, 492, and 214 were mutated to lysine. ( C ) Effects of downstream UPR signaling pathways after GRP78/BiP R470+492K and GRP78/BiP R214K plasmids transfected into G12D cells. * P < 0.01 by 1-way ANOVA with Tukey’s HSD test (mean ± SEM, n = 3). ( D ) Schematic diagram of arginine–mono-ADP-ribosylated GRP78/BiP involved in the regulation of ER homeostasis under the state of ER stress caused by KRAS mutation: ART1 catalyzes the arginine mono-ADP-ribosylation of GRP78/BiP to inactivate it and release the receptors bound to it. The downstream UPR signaling pathway is continuously activated to help maintain the homeostasis of the ER and the survival of tumor cells. ( E ) Schematic diagram of the effect of interference with arginine–mono-ADP-ribosylated GRP78/BiP on the UPR signaling pathway. Interfering with GRP78/BiP arginine mono-ADP-ribosylation modification can downregulate the expression of GRP78/BiP; limited GRP78/BiP can bind to unfolded protein and sensors PERK and IRE1α, inhibiting the IRE1α/XBP1/TFAF2/JNK pathway. The PERK/eIF2α/ATF4 signaling pathway further leads to the reduction of GRP78 expression, damages the ER stress regulation mechanism, increases the pressure of the internal environment, hinders the growth of tumor cells, and the cells tend to undergo apoptosis.

    Article Snippet: The following antibodies were used: ATF6 (catalog 3192), PERK (catalog 65880), IRE1α (catalog 3294), eIF2α (catalog 5324), p-eIF2α (catalog 3398) (all Cell Signaling Technology); GRP78/BiP (catalog sc-13539), HSC70 (catalog sc-24), Bcl2 (catalog sc-7382) (all Santa Cruz Biotechnology); p-IRE1α (catalog ab124945), XBP1u/s (catalog ab220783) (both Abcam); p-PERK (catalog DF7576, Affinity Biosciences); CHOP (catalog NB600-1335SS, Novus Biotechnology); ART1 (catalog A10103, Abclonal Technology Co., Ltd.); Caspase-3 (catalog WL02117), PARP/cleaved-PARP (catalog WL01932) (both Wanlei Biotechnology); β-actin (catalog 6609-1-Ig), Ki67 (catalog 27309-1-AP) (both Proteintech).

    Techniques: Protein-Protein interactions, Transfection, Mutagenesis, Modification, Expressing