Journal: eLife

Article Title: UBTD1 regulates ceramide balance and endolysosomal positioning to coordinate EGFR signaling

doi: 10.7554/eLife.68348

Figure Lengend Snippet: ( A–F ) DU145 cells were transfected for 48 hr with the indicated siRNA (siCTRLpool or siUBTD1pool). ( A,B ) Heatmap of all ( A ) lipids and ( B ) ceramide levels. The normalized expression of each lipid is shown in a scale range from blue to red. CER: ceramides; ChE: cholesterol ester; DG: diacylglycerols; LPC: lysophosphatidylcholines; MG: monoacylglycerols; PC: phosphatidylcholines; PE: phosphatidylethanolamines; PG: phosphatidylglycerol; PI: phosphatidylinositol; PS: phosphatidylserines; SM: sphingomyelins; TG: triglycerides; ZyE: zymosterols. ( C,D ) Mean intensities of positive-ion ( C ) and negative-ion ( D ) MS spectra from reflectron MALDI-TOF analyses. ( E ) Immunoblot and quantification of p-EGFR (Y1068 or Y1086) in the presence of different concentrations of GM3. p-EGFR levels were quantified by calculating the ratio between p-EGFR and EGFR, both normalized to loading control signal. Immunoblot of UBTD1 shows the level of siRNA depletion. ( F ) Representative cell growth curves measured by videomicroscopy (Incucyte, Essen Bioscience) in the presence of GM3. T = 0 corresponds to transfection time and the time window (40–60 hr) is presented. n ≥ 3 independent experiments. *p<0.05 **p<0.01, ***p<0.001; ****p<0.0001; ( C–F ) two-way ANOVA and Bonferroni’s multiple comparisons test; data are mean ± s.e.m. Figure 2—source data 1. Uncropped western blot for . Figure 2—source data 2. Row data for and for .

Article Snippet: Ganglioside GM3 (860058) was purchased from Avanti Polar Lipids.

Techniques: Transfection, Expressing, Western Blot, Control

Journal: eLife

Article Title: UBTD1 regulates ceramide balance and endolysosomal positioning to coordinate EGFR signaling

doi: 10.7554/eLife.68348

Figure Lengend Snippet: ( A–D ) DU145 cells were transfected for 48 hr with the indicated siRNA (control, siCTRLpool or UBTD1: siUBTD1pool). ( A ) Immunoblot and quantification of p62. The immunoblot of UBTD1 shows the level of siRNA depletion. ( B ) Immunoblots (up) and quantification (down) of p62 levels in cells treated with cycloheximide (CHX) at different time points. Immunoblot of UBTD1 shows the level of siRNA depletion. Hsp90 was used as a loading control. ( C ) Proximal ligation assay monitoring and quantification of p62 associated with RNF26. ( D ) Immunoblot and quantification of LC3 and p62 in cells treated or not with bafilomycin at different time points. The immunoblot of UBTD1 shows the level of siRNA depletion. ( E ) A proposed model for UBTD1 coordination of EGFR signaling and cell proliferation. EGF binding induces phosphorylation of EGFR and causes internalization of the EGF/EGFR complex. The complex is then transported by the endocytic system to be either recycled to the membrane or degraded by the lysosome. EGFR auto-phosphorylation is prevented by ganglioside 3 (GM3) (left). UBTD1 depletion decreases the ubiquitination of the ASAH1 leading to a decline of GM3. Decrease of GM3 induce EGFR self-phosphorylation and signaling. Depletion of UBTD1 also alters endolysosomal positioning and decreases p62/SQSTM1 ubiquitination leading to the inhibition of EGFR degradation. A decrease in EGFR degradation combined with an increase in auto-phosphorylation exacerbates EGFR signaling pathway and induces an EGFR-dependent cell proliferation (right). Scale bar = 10 µm. n ≥ 3 independent experiments; ns = non-significant; *p<0.05; ***p<0.001; ****p<0.0001; ( A,C ) two-tailed t-test; ( D ) Bonferroni’s multiple comparison test; data are mean ± s.e.m. Figure 6—figure supplement 1—source data 1. Uncropped western blot for .

Article Snippet: Ganglioside GM3 (860058) was purchased from Avanti Polar Lipids.

Techniques: Transfection, Control, Western Blot, Ligation, Binding Assay, Phospho-proteomics, Membrane, Ubiquitin Proteomics, Inhibition, Two Tailed Test, Comparison

Journal: Frontiers in Neurology

Article Title: Serial assessment of the novel biomarker MFGE8 in comparison with GFAP for predicting severity and outcome after traumatic brain injury

doi: 10.3389/fneur.2026.1750940

Figure Lengend Snippet: Serum biomarker concentrations over time after TBI. Serum levels of (A) MFGE8, (B) GFAP, and (C) the GFAP/MFGE8 ratio were measured in healthy controls and at admission, 24 h, 48 h, and 72 h after TBI. Data are presented as scatter-box plots (median with interquartile range) with log 10 -transformed concentrations.

Article Snippet: Serum levels of MFGE8 (E-EL-H2063) and GFAP (E-EL-H6093) were measured using a double-antibody sandwich ELISA (Elabscience Biotechnology Co., Ltd).

Techniques: Biomarker Discovery, Transformation Assay

Journal: Frontiers in Neurology

Article Title: Serial assessment of the novel biomarker MFGE8 in comparison with GFAP for predicting severity and outcome after traumatic brain injury

doi: 10.3389/fneur.2026.1750940

Figure Lengend Snippet: Temporal distribution of serum MFGE8 and GFAP levels, and the GFAP/MFGE8 ratio across TBI severity groups based on GCS scores. Serum levels of MFGE8, GFAP, and the GFAP/MFGE8 ratio in patients with (A) mild, (B) moderate, and (C) severe TBI (stratified by GCS) at admission and at 24, 48, and 72 h post-injury. Each row represents a distinct TBI severity group. Data are presented as scatter-box plots (median with interquartile range) with log 10 -transformed concentrations.

Article Snippet: Serum levels of MFGE8 (E-EL-H2063) and GFAP (E-EL-H6093) were measured using a double-antibody sandwich ELISA (Elabscience Biotechnology Co., Ltd).

Techniques: Transformation Assay

Journal: Frontiers in Neurology

Article Title: Serial assessment of the novel biomarker MFGE8 in comparison with GFAP for predicting severity and outcome after traumatic brain injury

doi: 10.3389/fneur.2026.1750940

Figure Lengend Snippet: Temporal profiles of serum MFGE8 and GFAP levels stratified by clinical outcomes. Line charts depict the temporal trajectories of serum (A) MFGE8 and (B) GFAP concentrations stratified by clinical outcomes, categorized as favorable alive (GOSE 5–8), unfavorable alive (GOSE 2–4), and death (GOSE 1), measured at admission and at 24, 48, and 72 h after traumatic brain injury. Data points represent median values, and shaded areas indicate the interquartile range (IQR).

Article Snippet: Serum levels of MFGE8 (E-EL-H2063) and GFAP (E-EL-H6093) were measured using a double-antibody sandwich ELISA (Elabscience Biotechnology Co., Ltd).

Techniques:

Journal: Frontiers in Neurology

Article Title: Serial assessment of the novel biomarker MFGE8 in comparison with GFAP for predicting severity and outcome after traumatic brain injury

doi: 10.3389/fneur.2026.1750940

Figure Lengend Snippet: ROC curve analysis of MFGE8, GFAP, and the GFAP/MFGE8 ratio for predicting 30-day mortality risk.

Article Snippet: Serum levels of MFGE8 (E-EL-H2063) and GFAP (E-EL-H6093) were measured using a double-antibody sandwich ELISA (Elabscience Biotechnology Co., Ltd).

Techniques: