Journal: Materials Today Bio

Article Title: Application of a novel myristoylproteomics approach identifies GLIPR2 as a key pro-ferroptotic substrate in non-small cell lung cancer

doi: 10.1016/j.mtbio.2026.102945

Figure Lengend Snippet: Augmented myristoylation is a feature and consequence of ferroptosis in sensitive cells. A. Workflow for metabolic labeling with YnMyr and detection of myristoylated proteins via CuAAC. B. Viability of Calu-1 cells pretreated with DMSO or 1 μM IMP-1088, then exposed to erastin or ML162 ± Fer-1. C. The impact of ferroptosis to N-myristoylation was detected by western blot. Calu-1 and H460 cells were first incubated with YnMyr for 18 h. For the final 8 h of YnMyr incubation, the cells were co-treated with 5 μM erastin in the presence or absence of 2 μM Fer-1. In a separate experiment, for the final 2 h of YnMyr incubation, the cells were co-treated with 1 μM RSL3 with or without 2 μM Fer-1. D. Samples were analyzed by TAMRA (top) or enriched by pull-down on streptavidin beads and analyzed by Western blot (bottom). The sample before pull-down (Input), pull-down sample (PD) and the supernatant from the pull-down (Spnt) were analyzed. C-SRC, PRKACA and CHCHD3 were enriched in the pull-down samples. GAPDH: loading control. E-F. The impact of ferroptosis to NMT1 or NMT2 was detected by western blot. Cells (Calu-1, E; H460, F) were treated with 5 μM erastin or 1 μM RSL3 for indicated time.Experiments in B were repeated in triplicate.

Article Snippet: IMP-1088 (#HY-112258), erastin (#HY-15763) were purchased from MedChemExpress (Monmouth, NJ, USA).

Techniques: Labeling, Western Blot, Incubation, Control

Journal: Materials Today Bio

Article Title: Application of a novel myristoylproteomics approach identifies GLIPR2 as a key pro-ferroptotic substrate in non-small cell lung cancer

doi: 10.1016/j.mtbio.2026.102945

Figure Lengend Snippet: Myristoylation-dependent ER localization is required for GLIPR2 to promote ferroptosis. A-B. Dose-response curves of GLIPR2 knockout Calu-1 cells with or without GLIPR2-WT or G2A overexpressed, following treated with erastin (A) or ML162 (B) for 24 h. Viability was assessed and normalized to control. C. Western blot analysis validating the expression of GLIPR2 (WT or G2A) in reconstituted GLIPR2-knockout Calu-1 cells. EV, empty vector. D. Western blot analysis of GLIPR2 myristoylation in GLIPR2-knockout Calu-1 cells reconstituted as indicated, and treated with or without 1 μM IMP-1088. E. Immunofluorescence analysis showing the subcellular localization of re-introduced wild-type GLIPR2 and the G2A mutant in GLIPR2-knockout Calu-1 cells. Scale bar, 20 μm. F-G. Dose-response curves of GLIPR2-knockout Calu-1 cells reconstituted with GLIPR2 (WT) or the indicated subcellular localization mutants, treated with erastin (F) or ML162 (G) for 24 h. H. Immunofluorescence analysis of the subcellular localization of the indicated GLIPR2 mutants in GLIPR2-knockout Calu-1 cells. Scale bar, 20 μm. I. Western blot analysis of the expression levels of GLIPR2 (WT) and the indicated mutants in GLIPR2-knockout Calu-1 cells. Data are mean ± SD; n = 3 (A–B and F-G).

Article Snippet: IMP-1088 (#HY-112258), erastin (#HY-15763) were purchased from MedChemExpress (Monmouth, NJ, USA).

Techniques: Knock-Out, Control, Western Blot, Expressing, Plasmid Preparation, Immunofluorescence, Mutagenesis

Journal: Materials Today Bio

Article Title: Application of a novel myristoylproteomics approach identifies GLIPR2 as a key pro-ferroptotic substrate in non-small cell lung cancer

doi: 10.1016/j.mtbio.2026.102945

Figure Lengend Snippet: Augmented myristoylation is a feature and consequence of ferroptosis in sensitive cells. A. Workflow for metabolic labeling with YnMyr and detection of myristoylated proteins via CuAAC. B. Viability of Calu-1 cells pretreated with DMSO or 1 μM IMP-1088, then exposed to erastin or ML162 ± Fer-1. C. The impact of ferroptosis to N-myristoylation was detected by western blot. Calu-1 and H460 cells were first incubated with YnMyr for 18 h. For the final 8 h of YnMyr incubation, the cells were co-treated with 5 μM erastin in the presence or absence of 2 μM Fer-1. In a separate experiment, for the final 2 h of YnMyr incubation, the cells were co-treated with 1 μM RSL3 with or without 2 μM Fer-1. D. Samples were analyzed by TAMRA (top) or enriched by pull-down on streptavidin beads and analyzed by Western blot (bottom). The sample before pull-down (Input), pull-down sample (PD) and the supernatant from the pull-down (Spnt) were analyzed. C-SRC, PRKACA and CHCHD3 were enriched in the pull-down samples. GAPDH: loading control. E-F. The impact of ferroptosis to NMT1 or NMT2 was detected by western blot. Cells (Calu-1, E; H460, F) were treated with 5 μM erastin or 1 μM RSL3 for indicated time.Experiments in B were repeated in triplicate.

Article Snippet: IMP-1088 (#HY-112258), erastin (#HY-15763) were purchased from MedChemExpress (Monmouth, NJ, USA).

Techniques: Labeling, Western Blot, Incubation, Control

Journal: Materials Today Bio

Article Title: Application of a novel myristoylproteomics approach identifies GLIPR2 as a key pro-ferroptotic substrate in non-small cell lung cancer

doi: 10.1016/j.mtbio.2026.102945

Figure Lengend Snippet: Comparative myristoylome profiling identifies candidates associated with ferroptosis sensitivity. A. Experimental strategy to identify N-myristoylated proteins with higher levels in ferroptosis-sensitive Calu-1 vs. H460 cells. B. Volcano plot for Calu-1 cells shows YnMyr incorporation versus significance in the presence of IMP-1088. Confidence categories: high (blue), medium (green; circle, rhombus, triangle), low (yellow), potential (purple), non-substrates (gray). C. Corresponding volcano plot for H460 cells. D. Comparison of YnMyr intensities between Calu-1 and H460 cells. Subtypes: Calu-1 > H460 (red), unique to Calu-1 (yellow), unique to H460 (green), H460 > Calu-1 (blue), common (purple), non-substrates (gray). E. Schematic for selecting myristoylated substrates linked to ferroptosis sensitivity. F. Viability of Calu-1 cells after siRNA knockdown of candidate genes and treatment with 0.04 μM ML162. Data are mean ± SD; n = 3 (F). Statistics: unpaired t-test (F). ∗p < 0.05, ∗∗p < 0.01, NS, not significant.

Article Snippet: IMP-1088 (#HY-112258), erastin (#HY-15763) were purchased from MedChemExpress (Monmouth, NJ, USA).

Techniques: Comparison, Knockdown

Journal: Materials Today Bio

Article Title: Application of a novel myristoylproteomics approach identifies GLIPR2 as a key pro-ferroptotic substrate in non-small cell lung cancer

doi: 10.1016/j.mtbio.2026.102945

Figure Lengend Snippet: Myristoylation-dependent ER localization is required for GLIPR2 to promote ferroptosis. A-B. Dose-response curves of GLIPR2 knockout Calu-1 cells with or without GLIPR2-WT or G2A overexpressed, following treated with erastin (A) or ML162 (B) for 24 h. Viability was assessed and normalized to control. C. Western blot analysis validating the expression of GLIPR2 (WT or G2A) in reconstituted GLIPR2-knockout Calu-1 cells. EV, empty vector. D. Western blot analysis of GLIPR2 myristoylation in GLIPR2-knockout Calu-1 cells reconstituted as indicated, and treated with or without 1 μM IMP-1088. E. Immunofluorescence analysis showing the subcellular localization of re-introduced wild-type GLIPR2 and the G2A mutant in GLIPR2-knockout Calu-1 cells. Scale bar, 20 μm. F-G. Dose-response curves of GLIPR2-knockout Calu-1 cells reconstituted with GLIPR2 (WT) or the indicated subcellular localization mutants, treated with erastin (F) or ML162 (G) for 24 h. H. Immunofluorescence analysis of the subcellular localization of the indicated GLIPR2 mutants in GLIPR2-knockout Calu-1 cells. Scale bar, 20 μm. I. Western blot analysis of the expression levels of GLIPR2 (WT) and the indicated mutants in GLIPR2-knockout Calu-1 cells. Data are mean ± SD; n = 3 (A–B and F-G).

Article Snippet: IMP-1088 (#HY-112258), erastin (#HY-15763) were purchased from MedChemExpress (Monmouth, NJ, USA).

Techniques: Knock-Out, Control, Western Blot, Expressing, Plasmid Preparation, Immunofluorescence, Mutagenesis