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Mettl18 deficiency induces unfolded protein response (UPR) activation and inflammatory cytokine expression. (A–D) Quantification of UPR pathway activation. Western blot analysis of whole pancreatic lysates from WT and KO mice using (A) anti-PERK (phospho-Thr980) and anti-PERK antibodies to assess PERK phosphorylation; (B) anti-EIF2 (phospho-Ser51) and anti-EIF2 antibodies to assess EIF2 phosphorylation; (C) anti-ATF4 antibody; and (D) anti-IRE1 (phospho-Ser724) and anti-IRE1 antibodies. Band intensities were quantified with ImageJ. 16-week-old; n = 6; mean ± SEM. Student's t -test: p∗∗<0.01, p∗<0.05. See also . (E) qPCR quantification of Xbp1s mRNA levels. Ct values were normalized to those obtained with primers detecting both Xbp1s and unspliced Xbp1 ( Xbp1u ) as described in . 16-week-old; n = 7; mean ± SEM. Student's t -test: n.s. > 0.1. See also . (F and G) qPCR quantification of mRNA expression of inflammatory cytokines, Il-1 β (E) and Il-6 (F). 16-week-old; n = 7; mean ± SEM. Student's t -test: p∗<0.05. (H) IL-1β protein levels were assessed by Western blotting of whole pancreatic lysates from WT and KO mice using an anti-IL-1β antibody. Quantification and statistical analyses were performed as in (A–D). (I) Schematic representation of UPR activation in WT and Mettl18 KO pancreas.

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: Mettl18 deficiency induces unfolded protein response (UPR) activation and inflammatory cytokine expression. (A–D) Quantification of UPR pathway activation. Western blot analysis of whole pancreatic lysates from WT and KO mice using (A) anti-PERK (phospho-Thr980) and anti-PERK antibodies to assess PERK phosphorylation; (B) anti-EIF2 (phospho-Ser51) and anti-EIF2 antibodies to assess EIF2 phosphorylation; (C) anti-ATF4 antibody; and (D) anti-IRE1 (phospho-Ser724) and anti-IRE1 antibodies. Band intensities were quantified with ImageJ. 16-week-old; n = 6; mean ± SEM. Student's t -test: p∗∗<0.01, p∗<0.05. See also . (E) qPCR quantification of Xbp1s mRNA levels. Ct values were normalized to those obtained with primers detecting both Xbp1s and unspliced Xbp1 ( Xbp1u ) as described in . 16-week-old; n = 7; mean ± SEM. Student's t -test: n.s. > 0.1. See also . (F and G) qPCR quantification of mRNA expression of inflammatory cytokines, Il-1 β (E) and Il-6 (F). 16-week-old; n = 7; mean ± SEM. Student's t -test: p∗<0.05. (H) IL-1β protein levels were assessed by Western blotting of whole pancreatic lysates from WT and KO mice using an anti-IL-1β antibody. Quantification and statistical analyses were performed as in (A–D). (I) Schematic representation of UPR activation in WT and Mettl18 KO pancreas.

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: Activation Assay, Expressing, Western Blot, Phospho-proteomics