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anti reg1  (Proteintech)


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    Proteintech anti reg1
    Proteomic alterations in the pancreas of Mettl18 -deficient mice. (A) Volcano plot of quantitative LC-MS/MS of the mouse pancreas. n = 3 per genotype. Proteins with |log2FC| > 0.5 and -Log 10 (P-value) > 4 are considered significantly changed. (B) Gene ontology (GO) analysis. A functional gene annotation of the 19 proteins (up) or 13 proteins (down) was performed with DAVID GO analysis (v6.8). Functional annotations against Biological Process, Molecular Function, and Cellular Component gene ontology with FDR less than 0.05 were shown. (C) Western blot of the pancreatic proteins from WT and KO mice. <t>Top;</t> <t>anti-Reg1</t> antibody, bottom; anti-β-actin antibody. (D) Quantification of Reg1 protein in the pancreas. 16-week-old; n = 6; mean ± SEM. Student's t -test: p∗∗ = 0.00423. (E) qPCR quantification of mRNA expression of Reg1 in the pancreas. 16-week-old; n = 7; mean ± SEM. Student's t -test: n.s. >0.1. (F) Distribution of Reg1-FLAG. WT and two independent Mettl18 KO clones (KO1 and KO2) of 266-6 cells were transfected with Reg1-FLAG. After 48 h, cells were fractionated into soluble and insoluble fractions, and Reg1-FLAG was analyzed by Western blotting. n = 3; mean ± SEM. Dunnett's test: p∗ <0.05. See also . (G) WT and Mettl18 KO cells were transfected with the plasmid for Reg1-FLAG. 48 h after transfection, cells were fixed, and Reg1-FLAG aggregation was observed under a microscope. Arrowhead indicates aggregation foci. Scale bar: 10 μm. (H) Quantification of 266-6 cells with Reg1-FLAG aggregation. Mean value of three independent experiments was shown. Dunnett's test: p∗∗ = 0.00195, p∗∗∗ = 0.00054. (I) Distribution of Reg1-FLAG in rescued cells. WT and KO1 cells were transfected with Reg1-FLAG together with or without METTL18-WT-HA or its catalytic mutant, METTL18-mut-HA. Forty-eight hours after, cells were fractionated into soluble and insoluble fractions, and Reg1-FLAG was analyzed by Western blotting. n = 3; mean ± SEM. Statistical significance was assessed using Dunnett's test with the KO group as the reference. P∗∗ <0.01. See also . (J) WT and KO1 cells transfected with Reg1-FLAG together with or without METTL18-WT-HA or METTL18-mut-HA were fixed 48 h after transfection, and FLAG signals were visualized by fluorescence microscopy. Scale bar: 10 μm. (K) Quantification of the Reg1-FLAG aggregation. n = 3; mean ± SEM. Dunnett's test: p∗∗ = 0.00498, p∗∗∗<0.0001.
    Anti Reg1, supplied by Proteintech, used in various techniques. Bioz Stars score: 91/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/anti+reg1/REG+Antibody/pmc12962108-259-19-24
    Average 91 stars, based on 3 article reviews
    anti reg1 - by Bioz Stars, 2026-09
    91/100 stars

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    1) Product Images from "METTL18 ensures pancreatic function by maintaining proper translation and proteostasis"

    Article Title: METTL18 ensures pancreatic function by maintaining proper translation and proteostasis

    Journal: Molecular Metabolism

    doi: 10.1016/j.molmet.2026.102337

    Proteomic alterations in the pancreas of Mettl18 -deficient mice. (A) Volcano plot of quantitative LC-MS/MS of the mouse pancreas. n = 3 per genotype. Proteins with |log2FC| > 0.5 and -Log 10 (P-value) > 4 are considered significantly changed. (B) Gene ontology (GO) analysis. A functional gene annotation of the 19 proteins (up) or 13 proteins (down) was performed with DAVID GO analysis (v6.8). Functional annotations against Biological Process, Molecular Function, and Cellular Component gene ontology with FDR less than 0.05 were shown. (C) Western blot of the pancreatic proteins from WT and KO mice. Top; anti-Reg1 antibody, bottom; anti-β-actin antibody. (D) Quantification of Reg1 protein in the pancreas. 16-week-old; n = 6; mean ± SEM. Student's t -test: p∗∗ = 0.00423. (E) qPCR quantification of mRNA expression of Reg1 in the pancreas. 16-week-old; n = 7; mean ± SEM. Student's t -test: n.s. >0.1. (F) Distribution of Reg1-FLAG. WT and two independent Mettl18 KO clones (KO1 and KO2) of 266-6 cells were transfected with Reg1-FLAG. After 48 h, cells were fractionated into soluble and insoluble fractions, and Reg1-FLAG was analyzed by Western blotting. n = 3; mean ± SEM. Dunnett's test: p∗ <0.05. See also . (G) WT and Mettl18 KO cells were transfected with the plasmid for Reg1-FLAG. 48 h after transfection, cells were fixed, and Reg1-FLAG aggregation was observed under a microscope. Arrowhead indicates aggregation foci. Scale bar: 10 μm. (H) Quantification of 266-6 cells with Reg1-FLAG aggregation. Mean value of three independent experiments was shown. Dunnett's test: p∗∗ = 0.00195, p∗∗∗ = 0.00054. (I) Distribution of Reg1-FLAG in rescued cells. WT and KO1 cells were transfected with Reg1-FLAG together with or without METTL18-WT-HA or its catalytic mutant, METTL18-mut-HA. Forty-eight hours after, cells were fractionated into soluble and insoluble fractions, and Reg1-FLAG was analyzed by Western blotting. n = 3; mean ± SEM. Statistical significance was assessed using Dunnett's test with the KO group as the reference. P∗∗ <0.01. See also . (J) WT and KO1 cells transfected with Reg1-FLAG together with or without METTL18-WT-HA or METTL18-mut-HA were fixed 48 h after transfection, and FLAG signals were visualized by fluorescence microscopy. Scale bar: 10 μm. (K) Quantification of the Reg1-FLAG aggregation. n = 3; mean ± SEM. Dunnett's test: p∗∗ = 0.00498, p∗∗∗<0.0001.
    Figure Legend Snippet: Proteomic alterations in the pancreas of Mettl18 -deficient mice. (A) Volcano plot of quantitative LC-MS/MS of the mouse pancreas. n = 3 per genotype. Proteins with |log2FC| > 0.5 and -Log 10 (P-value) > 4 are considered significantly changed. (B) Gene ontology (GO) analysis. A functional gene annotation of the 19 proteins (up) or 13 proteins (down) was performed with DAVID GO analysis (v6.8). Functional annotations against Biological Process, Molecular Function, and Cellular Component gene ontology with FDR less than 0.05 were shown. (C) Western blot of the pancreatic proteins from WT and KO mice. Top; anti-Reg1 antibody, bottom; anti-β-actin antibody. (D) Quantification of Reg1 protein in the pancreas. 16-week-old; n = 6; mean ± SEM. Student's t -test: p∗∗ = 0.00423. (E) qPCR quantification of mRNA expression of Reg1 in the pancreas. 16-week-old; n = 7; mean ± SEM. Student's t -test: n.s. >0.1. (F) Distribution of Reg1-FLAG. WT and two independent Mettl18 KO clones (KO1 and KO2) of 266-6 cells were transfected with Reg1-FLAG. After 48 h, cells were fractionated into soluble and insoluble fractions, and Reg1-FLAG was analyzed by Western blotting. n = 3; mean ± SEM. Dunnett's test: p∗ <0.05. See also . (G) WT and Mettl18 KO cells were transfected with the plasmid for Reg1-FLAG. 48 h after transfection, cells were fixed, and Reg1-FLAG aggregation was observed under a microscope. Arrowhead indicates aggregation foci. Scale bar: 10 μm. (H) Quantification of 266-6 cells with Reg1-FLAG aggregation. Mean value of three independent experiments was shown. Dunnett's test: p∗∗ = 0.00195, p∗∗∗ = 0.00054. (I) Distribution of Reg1-FLAG in rescued cells. WT and KO1 cells were transfected with Reg1-FLAG together with or without METTL18-WT-HA or its catalytic mutant, METTL18-mut-HA. Forty-eight hours after, cells were fractionated into soluble and insoluble fractions, and Reg1-FLAG was analyzed by Western blotting. n = 3; mean ± SEM. Statistical significance was assessed using Dunnett's test with the KO group as the reference. P∗∗ <0.01. See also . (J) WT and KO1 cells transfected with Reg1-FLAG together with or without METTL18-WT-HA or METTL18-mut-HA were fixed 48 h after transfection, and FLAG signals were visualized by fluorescence microscopy. Scale bar: 10 μm. (K) Quantification of the Reg1-FLAG aggregation. n = 3; mean ± SEM. Dunnett's test: p∗∗ = 0.00498, p∗∗∗<0.0001.

    Techniques Used: Liquid Chromatography with Mass Spectroscopy, Functional Assay, Western Blot, Expressing, Clone Assay, Transfection, Plasmid Preparation, Microscopy, Mutagenesis, Fluorescence

    Related Articles

    Liquid Chromatography with Mass Spectroscopy:

    Article Title: METTL18 ensures pancreatic function by maintaining proper translation and proteostasis
    Article Snippet: Rabbit polyclonal antibodies for Methyl-RPL3-H245 were obtained from ABclonal (cat#A18821).Rabbit polyclonal antibodies for Methyl-RPL3-H245 were obtained from ABclonal (cat#A18821).. Other antibodies used were as follows: anti-β-actin (clone 6D1, cat#M177-3; MBL); anti-phospho-PERK (Thr980) (clone G.305.4, cat#MA5-15033; Invitrogen); anti-RPL3 (cat#66130-1-Ig), anti-Reg1 (cat#15850-1-AP), and anti-PERK/EIF2AK3 (cat#24390-1-AP) (Proteintech Group); anti-IRE1 (cat#3294), anti-ATF4 (cat#11815), and anti-eIF5A (cat#20765) (Cell Signaling Technology); anti-phospho-IRE1 (cat#ab48187), anti-phospho-EIF2A (cat#ab32157), and anti-EIF2A (cat#ab5369) (Abcam); anti-IL-1β (cat#AF-401-NA; R&D Systems).. Unless otherwise indicated, all antibodies were used for Western blotting at a 1:1000 dilution in 5% skim milk in PBS containing 0.1% Tween-20, and incubated at 4 °C for 16 h with gentle agitation.Unless otherwise indicated, all antibodies were used for Western blotting at a 1:1000 dilution in 5% skim milk in PBS containing 0.1% Tween-20, and incubated at 4 °C for 16..

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    Functional Assay:

    Article Title: METTL18 ensures pancreatic function by maintaining proper translation and proteostasis
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    Western Blot:

    Article Title: METTL18 ensures pancreatic function by maintaining proper translation and proteostasis
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    Article Title: METTL18 Ensures Pancreatic Function by Maintaining Proper Translation and Proteostasis.
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    Expressing:

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    Article Title: METTL18 Ensures Pancreatic Function by Maintaining Proper Translation and Proteostasis.
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    Clone Assay:

    Article Title: METTL18 ensures pancreatic function by maintaining proper translation and proteostasis
    Article Snippet: Rabbit polyclonal antibodies for Methyl-RPL3-H245 were obtained from ABclonal (cat#A18821).Rabbit polyclonal antibodies for Methyl-RPL3-H245 were obtained from ABclonal (cat#A18821).. Other antibodies used were as follows: anti-β-actin (clone 6D1, cat#M177-3; MBL); anti-phospho-PERK (Thr980) (clone G.305.4, cat#MA5-15033; Invitrogen); anti-RPL3 (cat#66130-1-Ig), anti-Reg1 (cat#15850-1-AP), and anti-PERK/EIF2AK3 (cat#24390-1-AP) (Proteintech Group); anti-IRE1 (cat#3294), anti-ATF4 (cat#11815), and anti-eIF5A (cat#20765) (Cell Signaling Technology); anti-phospho-IRE1 (cat#ab48187), anti-phospho-EIF2A (cat#ab32157), and anti-EIF2A (cat#ab5369) (Abcam); anti-IL-1β (cat#AF-401-NA; R&D Systems).. Unless otherwise indicated, all antibodies were used for Western blotting at a 1:1000 dilution in 5% skim milk in PBS containing 0.1% Tween-20, and incubated at 4 °C for 16 h with gentle agitation.Unless otherwise indicated, all antibodies were used for Western blotting at a 1:1000 dilution in 5% skim milk in PBS containing 0.1% Tween-20, and incubated at 4 °C for 16..

    Article Title: METTL18 Ensures Pancreatic Function by Maintaining Proper Translation and Proteostasis.
    Article Snippet: Rabbit polyclonal antibodies for Methyl-RPL3-H245 were obtained from ABclonal 338 (cat#A18821).Rabbit polyclonal antibodies for Methyl-RPL3-H245 were obtained from ABclonal 338 (cat#A18821).. Other antibodies used were as follows: anti-β-actin (clone 6D1, cat#M177-3; MBL); 339 anti-phospho-PERK (Thr980) (clone G.305.4, cat#MA5-15033; Invitrogen); anti-RPL3 (cat#66130-1-340 Ig), anti-Reg1 (cat#15850-1-AP), and anti-PERK/EIF2AK3 (cat#24390-1-AP) (Proteintech Group); 341 anti-IRE1 (cat#3294), anti-ATF4 (cat#11815), and anti-eIF5A (cat#20765) (Cell Signaling 342 Technology); anti-phospho-IRE1 (cat#ab48187), anti-phospho-EIF2A (cat#ab32157), and anti-EIF2A 343 (cat#ab5369) (Abcam); anti-IL-1β (cat#AF-401-NA; R&D Systems).. Unless otherwise indicated, all 344 antibodies were used for Western blotting at a 1:1000 dilution in 5% skim milk in PBS containing 0.1% 345 Tween-20, and incubated at 4 °C for 16 h with gentle agitation.Unless otherwise indicated, all 344 antibodies were used for Western blotting at a 1:1000 dilution in 5% skim milk in PBS containing 0.1% 345 Tween-20, and incubated at 4..

    Transfection:

    Article Title: METTL18 ensures pancreatic function by maintaining proper translation and proteostasis
    Article Snippet: Rabbit polyclonal antibodies for Methyl-RPL3-H245 were obtained from ABclonal (cat#A18821).Rabbit polyclonal antibodies for Methyl-RPL3-H245 were obtained from ABclonal (cat#A18821).. Other antibodies used were as follows: anti-β-actin (clone 6D1, cat#M177-3; MBL); anti-phospho-PERK (Thr980) (clone G.305.4, cat#MA5-15033; Invitrogen); anti-RPL3 (cat#66130-1-Ig), anti-Reg1 (cat#15850-1-AP), and anti-PERK/EIF2AK3 (cat#24390-1-AP) (Proteintech Group); anti-IRE1 (cat#3294), anti-ATF4 (cat#11815), and anti-eIF5A (cat#20765) (Cell Signaling Technology); anti-phospho-IRE1 (cat#ab48187), anti-phospho-EIF2A (cat#ab32157), and anti-EIF2A (cat#ab5369) (Abcam); anti-IL-1β (cat#AF-401-NA; R&D Systems).. Unless otherwise indicated, all antibodies were used for Western blotting at a 1:1000 dilution in 5% skim milk in PBS containing 0.1% Tween-20, and incubated at 4 °C for 16 h with gentle agitation.Unless otherwise indicated, all antibodies were used for Western blotting at a 1:1000 dilution in 5% skim milk in PBS containing 0.1% Tween-20, and incubated at 4 °C for 16..

    Article Title: METTL18 Ensures Pancreatic Function by Maintaining Proper Translation and Proteostasis.
    Article Snippet: Rabbit polyclonal antibodies for Methyl-RPL3-H245 were obtained from ABclonal 338 (cat#A18821).Rabbit polyclonal antibodies for Methyl-RPL3-H245 were obtained from ABclonal 338 (cat#A18821).. Other antibodies used were as follows: anti-β-actin (clone 6D1, cat#M177-3; MBL); 339 anti-phospho-PERK (Thr980) (clone G.305.4, cat#MA5-15033; Invitrogen); anti-RPL3 (cat#66130-1-340 Ig), anti-Reg1 (cat#15850-1-AP), and anti-PERK/EIF2AK3 (cat#24390-1-AP) (Proteintech Group); 341 anti-IRE1 (cat#3294), anti-ATF4 (cat#11815), and anti-eIF5A (cat#20765) (Cell Signaling 342 Technology); anti-phospho-IRE1 (cat#ab48187), anti-phospho-EIF2A (cat#ab32157), and anti-EIF2A 343 (cat#ab5369) (Abcam); anti-IL-1β (cat#AF-401-NA; R&D Systems).. Unless otherwise indicated, all 344 antibodies were used for Western blotting at a 1:1000 dilution in 5% skim milk in PBS containing 0.1% 345 Tween-20, and incubated at 4 °C for 16 h with gentle agitation.Unless otherwise indicated, all 344 antibodies were used for Western blotting at a 1:1000 dilution in 5% skim milk in PBS containing 0.1% 345 Tween-20, and incubated at 4..

    Plasmid Preparation:

    Article Title: METTL18 ensures pancreatic function by maintaining proper translation and proteostasis
    Article Snippet: Rabbit polyclonal antibodies for Methyl-RPL3-H245 were obtained from ABclonal (cat#A18821).Rabbit polyclonal antibodies for Methyl-RPL3-H245 were obtained from ABclonal (cat#A18821).. Other antibodies used were as follows: anti-β-actin (clone 6D1, cat#M177-3; MBL); anti-phospho-PERK (Thr980) (clone G.305.4, cat#MA5-15033; Invitrogen); anti-RPL3 (cat#66130-1-Ig), anti-Reg1 (cat#15850-1-AP), and anti-PERK/EIF2AK3 (cat#24390-1-AP) (Proteintech Group); anti-IRE1 (cat#3294), anti-ATF4 (cat#11815), and anti-eIF5A (cat#20765) (Cell Signaling Technology); anti-phospho-IRE1 (cat#ab48187), anti-phospho-EIF2A (cat#ab32157), and anti-EIF2A (cat#ab5369) (Abcam); anti-IL-1β (cat#AF-401-NA; R&D Systems).. Unless otherwise indicated, all antibodies were used for Western blotting at a 1:1000 dilution in 5% skim milk in PBS containing 0.1% Tween-20, and incubated at 4 °C for 16 h with gentle agitation.Unless otherwise indicated, all antibodies were used for Western blotting at a 1:1000 dilution in 5% skim milk in PBS containing 0.1% Tween-20, and incubated at 4 °C for 16..

    Article Title: METTL18 Ensures Pancreatic Function by Maintaining Proper Translation and Proteostasis.
    Article Snippet: Rabbit polyclonal antibodies for Methyl-RPL3-H245 were obtained from ABclonal 338 (cat#A18821).Rabbit polyclonal antibodies for Methyl-RPL3-H245 were obtained from ABclonal 338 (cat#A18821).. Other antibodies used were as follows: anti-β-actin (clone 6D1, cat#M177-3; MBL); 339 anti-phospho-PERK (Thr980) (clone G.305.4, cat#MA5-15033; Invitrogen); anti-RPL3 (cat#66130-1-340 Ig), anti-Reg1 (cat#15850-1-AP), and anti-PERK/EIF2AK3 (cat#24390-1-AP) (Proteintech Group); 341 anti-IRE1 (cat#3294), anti-ATF4 (cat#11815), and anti-eIF5A (cat#20765) (Cell Signaling 342 Technology); anti-phospho-IRE1 (cat#ab48187), anti-phospho-EIF2A (cat#ab32157), and anti-EIF2A 343 (cat#ab5369) (Abcam); anti-IL-1β (cat#AF-401-NA; R&D Systems).. Unless otherwise indicated, all 344 antibodies were used for Western blotting at a 1:1000 dilution in 5% skim milk in PBS containing 0.1% 345 Tween-20, and incubated at 4 °C for 16 h with gentle agitation.Unless otherwise indicated, all 344 antibodies were used for Western blotting at a 1:1000 dilution in 5% skim milk in PBS containing 0.1% 345 Tween-20, and incubated at 4..

    Microscopy:

    Article Title: METTL18 ensures pancreatic function by maintaining proper translation and proteostasis
    Article Snippet: Rabbit polyclonal antibodies for Methyl-RPL3-H245 were obtained from ABclonal (cat#A18821).Rabbit polyclonal antibodies for Methyl-RPL3-H245 were obtained from ABclonal (cat#A18821).. Other antibodies used were as follows: anti-β-actin (clone 6D1, cat#M177-3; MBL); anti-phospho-PERK (Thr980) (clone G.305.4, cat#MA5-15033; Invitrogen); anti-RPL3 (cat#66130-1-Ig), anti-Reg1 (cat#15850-1-AP), and anti-PERK/EIF2AK3 (cat#24390-1-AP) (Proteintech Group); anti-IRE1 (cat#3294), anti-ATF4 (cat#11815), and anti-eIF5A (cat#20765) (Cell Signaling Technology); anti-phospho-IRE1 (cat#ab48187), anti-phospho-EIF2A (cat#ab32157), and anti-EIF2A (cat#ab5369) (Abcam); anti-IL-1β (cat#AF-401-NA; R&D Systems).. Unless otherwise indicated, all antibodies were used for Western blotting at a 1:1000 dilution in 5% skim milk in PBS containing 0.1% Tween-20, and incubated at 4 °C for 16 h with gentle agitation.Unless otherwise indicated, all antibodies were used for Western blotting at a 1:1000 dilution in 5% skim milk in PBS containing 0.1% Tween-20, and incubated at 4 °C for 16..

    Article Title: METTL18 Ensures Pancreatic Function by Maintaining Proper Translation and Proteostasis.
    Article Snippet: Rabbit polyclonal antibodies for Methyl-RPL3-H245 were obtained from ABclonal 338 (cat#A18821).Rabbit polyclonal antibodies for Methyl-RPL3-H245 were obtained from ABclonal 338 (cat#A18821).. Other antibodies used were as follows: anti-β-actin (clone 6D1, cat#M177-3; MBL); 339 anti-phospho-PERK (Thr980) (clone G.305.4, cat#MA5-15033; Invitrogen); anti-RPL3 (cat#66130-1-340 Ig), anti-Reg1 (cat#15850-1-AP), and anti-PERK/EIF2AK3 (cat#24390-1-AP) (Proteintech Group); 341 anti-IRE1 (cat#3294), anti-ATF4 (cat#11815), and anti-eIF5A (cat#20765) (Cell Signaling 342 Technology); anti-phospho-IRE1 (cat#ab48187), anti-phospho-EIF2A (cat#ab32157), and anti-EIF2A 343 (cat#ab5369) (Abcam); anti-IL-1β (cat#AF-401-NA; R&D Systems).. Unless otherwise indicated, all 344 antibodies were used for Western blotting at a 1:1000 dilution in 5% skim milk in PBS containing 0.1% 345 Tween-20, and incubated at 4 °C for 16 h with gentle agitation.Unless otherwise indicated, all 344 antibodies were used for Western blotting at a 1:1000 dilution in 5% skim milk in PBS containing 0.1% 345 Tween-20, and incubated at 4..

    Mutagenesis:

    Article Title: METTL18 ensures pancreatic function by maintaining proper translation and proteostasis
    Article Snippet: Rabbit polyclonal antibodies for Methyl-RPL3-H245 were obtained from ABclonal (cat#A18821).Rabbit polyclonal antibodies for Methyl-RPL3-H245 were obtained from ABclonal (cat#A18821).. Other antibodies used were as follows: anti-β-actin (clone 6D1, cat#M177-3; MBL); anti-phospho-PERK (Thr980) (clone G.305.4, cat#MA5-15033; Invitrogen); anti-RPL3 (cat#66130-1-Ig), anti-Reg1 (cat#15850-1-AP), and anti-PERK/EIF2AK3 (cat#24390-1-AP) (Proteintech Group); anti-IRE1 (cat#3294), anti-ATF4 (cat#11815), and anti-eIF5A (cat#20765) (Cell Signaling Technology); anti-phospho-IRE1 (cat#ab48187), anti-phospho-EIF2A (cat#ab32157), and anti-EIF2A (cat#ab5369) (Abcam); anti-IL-1β (cat#AF-401-NA; R&D Systems).. Unless otherwise indicated, all antibodies were used for Western blotting at a 1:1000 dilution in 5% skim milk in PBS containing 0.1% Tween-20, and incubated at 4 °C for 16 h with gentle agitation.Unless otherwise indicated, all antibodies were used for Western blotting at a 1:1000 dilution in 5% skim milk in PBS containing 0.1% Tween-20, and incubated at 4 °C for 16..

    Article Title: METTL18 Ensures Pancreatic Function by Maintaining Proper Translation and Proteostasis.
    Article Snippet: Rabbit polyclonal antibodies for Methyl-RPL3-H245 were obtained from ABclonal 338 (cat#A18821).Rabbit polyclonal antibodies for Methyl-RPL3-H245 were obtained from ABclonal 338 (cat#A18821).. Other antibodies used were as follows: anti-β-actin (clone 6D1, cat#M177-3; MBL); 339 anti-phospho-PERK (Thr980) (clone G.305.4, cat#MA5-15033; Invitrogen); anti-RPL3 (cat#66130-1-340 Ig), anti-Reg1 (cat#15850-1-AP), and anti-PERK/EIF2AK3 (cat#24390-1-AP) (Proteintech Group); 341 anti-IRE1 (cat#3294), anti-ATF4 (cat#11815), and anti-eIF5A (cat#20765) (Cell Signaling 342 Technology); anti-phospho-IRE1 (cat#ab48187), anti-phospho-EIF2A (cat#ab32157), and anti-EIF2A 343 (cat#ab5369) (Abcam); anti-IL-1β (cat#AF-401-NA; R&D Systems).. Unless otherwise indicated, all 344 antibodies were used for Western blotting at a 1:1000 dilution in 5% skim milk in PBS containing 0.1% 345 Tween-20, and incubated at 4 °C for 16 h with gentle agitation.Unless otherwise indicated, all 344 antibodies were used for Western blotting at a 1:1000 dilution in 5% skim milk in PBS containing 0.1% 345 Tween-20, and incubated at 4..

    Fluorescence:

    Article Title: METTL18 ensures pancreatic function by maintaining proper translation and proteostasis
    Article Snippet: Rabbit polyclonal antibodies for Methyl-RPL3-H245 were obtained from ABclonal (cat#A18821).Rabbit polyclonal antibodies for Methyl-RPL3-H245 were obtained from ABclonal (cat#A18821).. Other antibodies used were as follows: anti-β-actin (clone 6D1, cat#M177-3; MBL); anti-phospho-PERK (Thr980) (clone G.305.4, cat#MA5-15033; Invitrogen); anti-RPL3 (cat#66130-1-Ig), anti-Reg1 (cat#15850-1-AP), and anti-PERK/EIF2AK3 (cat#24390-1-AP) (Proteintech Group); anti-IRE1 (cat#3294), anti-ATF4 (cat#11815), and anti-eIF5A (cat#20765) (Cell Signaling Technology); anti-phospho-IRE1 (cat#ab48187), anti-phospho-EIF2A (cat#ab32157), and anti-EIF2A (cat#ab5369) (Abcam); anti-IL-1β (cat#AF-401-NA; R&D Systems).. Unless otherwise indicated, all antibodies were used for Western blotting at a 1:1000 dilution in 5% skim milk in PBS containing 0.1% Tween-20, and incubated at 4 °C for 16 h with gentle agitation.Unless otherwise indicated, all antibodies were used for Western blotting at a 1:1000 dilution in 5% skim milk in PBS containing 0.1% Tween-20, and incubated at 4 °C for 16..

    Article Title: METTL18 Ensures Pancreatic Function by Maintaining Proper Translation and Proteostasis.
    Article Snippet: Rabbit polyclonal antibodies for Methyl-RPL3-H245 were obtained from ABclonal 338 (cat#A18821).Rabbit polyclonal antibodies for Methyl-RPL3-H245 were obtained from ABclonal 338 (cat#A18821).. Other antibodies used were as follows: anti-β-actin (clone 6D1, cat#M177-3; MBL); 339 anti-phospho-PERK (Thr980) (clone G.305.4, cat#MA5-15033; Invitrogen); anti-RPL3 (cat#66130-1-340 Ig), anti-Reg1 (cat#15850-1-AP), and anti-PERK/EIF2AK3 (cat#24390-1-AP) (Proteintech Group); 341 anti-IRE1 (cat#3294), anti-ATF4 (cat#11815), and anti-eIF5A (cat#20765) (Cell Signaling 342 Technology); anti-phospho-IRE1 (cat#ab48187), anti-phospho-EIF2A (cat#ab32157), and anti-EIF2A 343 (cat#ab5369) (Abcam); anti-IL-1β (cat#AF-401-NA; R&D Systems).. Unless otherwise indicated, all 344 antibodies were used for Western blotting at a 1:1000 dilution in 5% skim milk in PBS containing 0.1% 345 Tween-20, and incubated at 4 °C for 16 h with gentle agitation.Unless otherwise indicated, all 344 antibodies were used for Western blotting at a 1:1000 dilution in 5% skim milk in PBS containing 0.1% 345 Tween-20, and incubated at 4..



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    Journal: Molecular Metabolism

    Article Title: METTL18 ensures pancreatic function by maintaining proper translation and proteostasis

    doi: 10.1016/j.molmet.2026.102337

    Figure Lengend Snippet: Proteomic alterations in the pancreas of Mettl18 -deficient mice. (A) Volcano plot of quantitative LC-MS/MS of the mouse pancreas. n = 3 per genotype. Proteins with |log2FC| > 0.5 and -Log 10 (P-value) > 4 are considered significantly changed. (B) Gene ontology (GO) analysis. A functional gene annotation of the 19 proteins (up) or 13 proteins (down) was performed with DAVID GO analysis (v6.8). Functional annotations against Biological Process, Molecular Function, and Cellular Component gene ontology with FDR less than 0.05 were shown. (C) Western blot of the pancreatic proteins from WT and KO mice. Top; anti-Reg1 antibody, bottom; anti-β-actin antibody. (D) Quantification of Reg1 protein in the pancreas. 16-week-old; n = 6; mean ± SEM. Student's t -test: p∗∗ = 0.00423. (E) qPCR quantification of mRNA expression of Reg1 in the pancreas. 16-week-old; n = 7; mean ± SEM. Student's t -test: n.s. >0.1. (F) Distribution of Reg1-FLAG. WT and two independent Mettl18 KO clones (KO1 and KO2) of 266-6 cells were transfected with Reg1-FLAG. After 48 h, cells were fractionated into soluble and insoluble fractions, and Reg1-FLAG was analyzed by Western blotting. n = 3; mean ± SEM. Dunnett's test: p∗ <0.05. See also . (G) WT and Mettl18 KO cells were transfected with the plasmid for Reg1-FLAG. 48 h after transfection, cells were fixed, and Reg1-FLAG aggregation was observed under a microscope. Arrowhead indicates aggregation foci. Scale bar: 10 μm. (H) Quantification of 266-6 cells with Reg1-FLAG aggregation. Mean value of three independent experiments was shown. Dunnett's test: p∗∗ = 0.00195, p∗∗∗ = 0.00054. (I) Distribution of Reg1-FLAG in rescued cells. WT and KO1 cells were transfected with Reg1-FLAG together with or without METTL18-WT-HA or its catalytic mutant, METTL18-mut-HA. Forty-eight hours after, cells were fractionated into soluble and insoluble fractions, and Reg1-FLAG was analyzed by Western blotting. n = 3; mean ± SEM. Statistical significance was assessed using Dunnett's test with the KO group as the reference. P∗∗ <0.01. See also . (J) WT and KO1 cells transfected with Reg1-FLAG together with or without METTL18-WT-HA or METTL18-mut-HA were fixed 48 h after transfection, and FLAG signals were visualized by fluorescence microscopy. Scale bar: 10 μm. (K) Quantification of the Reg1-FLAG aggregation. n = 3; mean ± SEM. Dunnett's test: p∗∗ = 0.00498, p∗∗∗<0.0001.

    Article Snippet: Other antibodies used were as follows: anti-β-actin (clone 6D1, cat#M177-3; MBL); anti-phospho-PERK (Thr980) (clone G.305.4, cat#MA5-15033; Invitrogen); anti-RPL3 (cat#66130-1-Ig), anti-Reg1 (cat#15850-1-AP), and anti-PERK/EIF2AK3 (cat#24390-1-AP) (Proteintech Group); anti-IRE1 (cat#3294), anti-ATF4 (cat#11815), and anti-eIF5A (cat#20765) (Cell Signaling Technology); anti-phospho-IRE1 (cat#ab48187), anti-phospho-EIF2A (cat#ab32157), and anti-EIF2A (cat#ab5369) (Abcam); anti-IL-1β (cat#AF-401-NA; R&D Systems).

    Techniques: Liquid Chromatography with Mass Spectroscopy, Functional Assay, Western Blot, Expressing, Clone Assay, Transfection, Plasmid Preparation, Microscopy, Mutagenesis, Fluorescence

    shRNA-induced knockdown of REG3A potently inhibits breast cancer cell proliferation and migration. The mRNA and protein expression of REG3A and REG1 in the stable pBC-1 primary breast cancer cells with the applied REG3A shRNA (“shREG3A-Sq2/3/6”, with non-overlapping sequences), the scramble control shRNA (“shC”), or in the parental control cells (“Ctrl”) was shown ( A and B ). Cells were further cultivated for indicated hours, cell proliferation (by measuring nuclear EdU incorporation, C ), cell migration ( D ) and invasion ( E ) were tested. The pBC-2 primary cancer cells, MCF-7 and MDA-231 lines, the primary mammary epithelial cells (pMEC) or established MCF-10A epithelial cells with shC and shREG3A-Sq6 were formed, and REG3A mRNA expression tested ( F and I ). Cells were further cultivated for designated hours, cell proliferation ( G and J ) and in vitro cell migration ( H ) were examined similarly. Values were mean ± standard deviation (SD, n = 5). * P < 0.05 versus “shC” group. “N. S.” stands for non-statistical difference ( P > 0.05). Experiments were repeated five times and similar results were obtained each time. Scale bar = 100 μm

    Journal: Breast Cancer Research : BCR

    Article Title: Increased expression of REG3A promotes tumorigenic behavior in triple negative breast cancer cells

    doi: 10.1186/s13058-024-01845-2

    Figure Lengend Snippet: shRNA-induced knockdown of REG3A potently inhibits breast cancer cell proliferation and migration. The mRNA and protein expression of REG3A and REG1 in the stable pBC-1 primary breast cancer cells with the applied REG3A shRNA (“shREG3A-Sq2/3/6”, with non-overlapping sequences), the scramble control shRNA (“shC”), or in the parental control cells (“Ctrl”) was shown ( A and B ). Cells were further cultivated for indicated hours, cell proliferation (by measuring nuclear EdU incorporation, C ), cell migration ( D ) and invasion ( E ) were tested. The pBC-2 primary cancer cells, MCF-7 and MDA-231 lines, the primary mammary epithelial cells (pMEC) or established MCF-10A epithelial cells with shC and shREG3A-Sq6 were formed, and REG3A mRNA expression tested ( F and I ). Cells were further cultivated for designated hours, cell proliferation ( G and J ) and in vitro cell migration ( H ) were examined similarly. Values were mean ± standard deviation (SD, n = 5). * P < 0.05 versus “shC” group. “N. S.” stands for non-statistical difference ( P > 0.05). Experiments were repeated five times and similar results were obtained each time. Scale bar = 100 μm

    Article Snippet: The anti-REG3A antibody (ab202057) and anti-REG1 antibody (ab47099) were obtained from Abcam (Shanghai, China).

    Techniques: shRNA, Migration, Expressing, In Vitro, Standard Deviation

    Figure 1. Reg1-3 genes were not necessary for pancreatic development and general growth in mice. (a) Schema of Reg1-3 genes location of human and mouse. We deleted about 100 kb, including no other genes. (b) 4-week-old and 8-week-old body weight among Reg+/+, Reg+/− and Reg−/− male and female mice. Bars indicate mean ± SEM (n = 6–12 mice). Statistical analysis was performed by one-way ANOVA with Tukey’s multiple comparison tests. (c) Representative hematoxylin and eosin (H&E) staining of pancreas tissue sections isolated from Reg+/+ and Reg−/− mice at 8-week-old during normal feeding (n = 4 mice). Scale bars: 100 μm.

    Journal: Scientific reports

    Article Title: Reg family proteins contribute to inflammation and pancreatic stellate cells activation in chronic pancreatitis.

    doi: 10.1038/s41598-023-39178-3

    Figure Lengend Snippet: Figure 1. Reg1-3 genes were not necessary for pancreatic development and general growth in mice. (a) Schema of Reg1-3 genes location of human and mouse. We deleted about 100 kb, including no other genes. (b) 4-week-old and 8-week-old body weight among Reg+/+, Reg+/− and Reg−/− male and female mice. Bars indicate mean ± SEM (n = 6–12 mice). Statistical analysis was performed by one-way ANOVA with Tukey’s multiple comparison tests. (c) Representative hematoxylin and eosin (H&E) staining of pancreas tissue sections isolated from Reg+/+ and Reg−/− mice at 8-week-old during normal feeding (n = 4 mice). Scale bars: 100 μm.

    Article Snippet: Antibodies used in this study were against Amylase (sc-12821, Santa Cruz), alpha smooth muscle Actin (αSMA, ab5694, abcam), Desmin (ab32362, abcam), Reg1 (AF1657, R&D), Reg2(AF1658, R&D), Reg3b(AF1660, R&D), Akt (#9272, CST),p44/42 MAPK (Erk1/2, #9102, CST) ; Horseradish peroxidase-conjugated donkey anti-rabbit IgG (NA9340, GE Healthcare Life Sciences), rabbit anti-goat IgG(AP106P, CHEMICON), rabbit-anti-sheep IgG (#313-035-003, Jackson Immuno Research Laboratories).

    Techniques: Comparison, Staining, Isolation

    Figure 5. Reg family proteins stimulated PSCs activation contributing to fibrosis in chronic pancreatitis. (a) Immunofluorescence analysis of alpha-smooth muscle actin (αSMA, an activated PSCs marker, red) in the pancreas sections of indicated genotypes at 2 weeks, and 4 weeks (n = 3–4 mice). Nuclei were counterstained by DAPI (blue). (b) The percentage of epithelial cells with αSMA positive signals. Results represent mean ± SEM (n = 3–4 mice). Statistical analysis was performed by one-way ANOVA with Tukey’s multiple comparison tests among four groups. **P < 0.01 (c) Cropped images of western blot gels of Amylase, αSMA, Desmin, Reg1, Reg2, Reg3b of the pancreas of indicated genotypes at 0.5 day after birth(P0.5), 1w and 4w, p44/42 MAPK (ERK 1/2) and Akt were used as the loading control (n = 3 mice). The samples derived from the same experiment and gels were processed in parallel. Images of the entire gels are presented in Supplementary Fig. 5–7. The right panel showed the respective densitometric quantification analysis of the relative intensity of αSMA. Densitometric quantification analysis of other proteins is presented in Supplementary Fig. 4.Results represent mean ± SEM (n = 3 mice). Statistical analysis was performed by one-way ANOVA with Tukey’s multiple comparison tests among four groups. ns: not significant; #P < 0.0001.

    Journal: Scientific reports

    Article Title: Reg family proteins contribute to inflammation and pancreatic stellate cells activation in chronic pancreatitis.

    doi: 10.1038/s41598-023-39178-3

    Figure Lengend Snippet: Figure 5. Reg family proteins stimulated PSCs activation contributing to fibrosis in chronic pancreatitis. (a) Immunofluorescence analysis of alpha-smooth muscle actin (αSMA, an activated PSCs marker, red) in the pancreas sections of indicated genotypes at 2 weeks, and 4 weeks (n = 3–4 mice). Nuclei were counterstained by DAPI (blue). (b) The percentage of epithelial cells with αSMA positive signals. Results represent mean ± SEM (n = 3–4 mice). Statistical analysis was performed by one-way ANOVA with Tukey’s multiple comparison tests among four groups. **P < 0.01 (c) Cropped images of western blot gels of Amylase, αSMA, Desmin, Reg1, Reg2, Reg3b of the pancreas of indicated genotypes at 0.5 day after birth(P0.5), 1w and 4w, p44/42 MAPK (ERK 1/2) and Akt were used as the loading control (n = 3 mice). The samples derived from the same experiment and gels were processed in parallel. Images of the entire gels are presented in Supplementary Fig. 5–7. The right panel showed the respective densitometric quantification analysis of the relative intensity of αSMA. Densitometric quantification analysis of other proteins is presented in Supplementary Fig. 4.Results represent mean ± SEM (n = 3 mice). Statistical analysis was performed by one-way ANOVA with Tukey’s multiple comparison tests among four groups. ns: not significant; #P < 0.0001.

    Article Snippet: Antibodies used in this study were against Amylase (sc-12821, Santa Cruz), alpha smooth muscle Actin (αSMA, ab5694, abcam), Desmin (ab32362, abcam), Reg1 (AF1657, R&D), Reg2(AF1658, R&D), Reg3b(AF1660, R&D), Akt (#9272, CST),p44/42 MAPK (Erk1/2, #9102, CST) ; Horseradish peroxidase-conjugated donkey anti-rabbit IgG (NA9340, GE Healthcare Life Sciences), rabbit anti-goat IgG(AP106P, CHEMICON), rabbit-anti-sheep IgG (#313-035-003, Jackson Immuno Research Laboratories).

    Techniques: Activation Assay, Immunofluorescence, Marker, Comparison, Western Blot, Control, Derivative Assay

    Figure 6. Reg deficiency led to the remission of chronic pancreatitis. (a) Representative pancreatic tissue section from mice of the indicated genotype at 8 weeks was stained with H&E or Azan (for collagen, dark blue) and Immunofluorescence staining of alpha-smooth muscle actin (αSMA, an activated PSCs marker, red) (n = 5 mice). Scale bars: H&E, 50 μm; Azan, 100 μm (b) Pancreatic mRNA expression of proinflammatory cytokines interleukin 6(Il6), tumor necrosis factor a (Tnfa) and Il1b of indicated genotypes were assessed by qRT-PCR. The expression amounts of each gene were calculated relative to those of ribosomal protein S3 (Rps3) with the fold change to CP model_Reg+/+ mice. Mean ± SEM (n = 5 mice). Statistical analysis was performed by two-tailed unpaired Student t-test between two groups. *P < 0.05 (c) Cropped images of western blot gels of Amylase, αSMA and Reg1 of the pancreas of indicated genotypes at 8 weeks and Akt was used as the loading control (n = 5 mice). The samples derived from the same experiment and gels were processed in parallel. Images of the entire gels and respective densitometric quantification analysis of the relative intensity of protein are presented in Supplementary Fig. 4,8.

    Journal: Scientific reports

    Article Title: Reg family proteins contribute to inflammation and pancreatic stellate cells activation in chronic pancreatitis.

    doi: 10.1038/s41598-023-39178-3

    Figure Lengend Snippet: Figure 6. Reg deficiency led to the remission of chronic pancreatitis. (a) Representative pancreatic tissue section from mice of the indicated genotype at 8 weeks was stained with H&E or Azan (for collagen, dark blue) and Immunofluorescence staining of alpha-smooth muscle actin (αSMA, an activated PSCs marker, red) (n = 5 mice). Scale bars: H&E, 50 μm; Azan, 100 μm (b) Pancreatic mRNA expression of proinflammatory cytokines interleukin 6(Il6), tumor necrosis factor a (Tnfa) and Il1b of indicated genotypes were assessed by qRT-PCR. The expression amounts of each gene were calculated relative to those of ribosomal protein S3 (Rps3) with the fold change to CP model_Reg+/+ mice. Mean ± SEM (n = 5 mice). Statistical analysis was performed by two-tailed unpaired Student t-test between two groups. *P < 0.05 (c) Cropped images of western blot gels of Amylase, αSMA and Reg1 of the pancreas of indicated genotypes at 8 weeks and Akt was used as the loading control (n = 5 mice). The samples derived from the same experiment and gels were processed in parallel. Images of the entire gels and respective densitometric quantification analysis of the relative intensity of protein are presented in Supplementary Fig. 4,8.

    Article Snippet: Antibodies used in this study were against Amylase (sc-12821, Santa Cruz), alpha smooth muscle Actin (αSMA, ab5694, abcam), Desmin (ab32362, abcam), Reg1 (AF1657, R&D), Reg2(AF1658, R&D), Reg3b(AF1660, R&D), Akt (#9272, CST),p44/42 MAPK (Erk1/2, #9102, CST) ; Horseradish peroxidase-conjugated donkey anti-rabbit IgG (NA9340, GE Healthcare Life Sciences), rabbit anti-goat IgG(AP106P, CHEMICON), rabbit-anti-sheep IgG (#313-035-003, Jackson Immuno Research Laboratories).

    Techniques: Staining, Immunofluorescence, Marker, Expressing, Quantitative RT-PCR, Two Tailed Test, Western Blot, Control, Derivative Assay