Journal: iScience

Article Title: The microcephaly-associated protein YIPF5 differentially regulates ER export

doi: 10.1016/j.isci.2026.114791

Figure Lengend Snippet: YIPF5 regulates the secretome composition (A) Schematic representation of the experimental workflow for analyzing the glycoprotein secretome. MCF10A cells were metabolically labeled with the clickable sugar analog ManNAz, followed by a biotin-azide click reaction to tag glycoproteins ( n = 4). Secreted glycoproteins were concentrated from the culture medium, selectively purified, and identified using mass spectrometry. The image was created with BioRender.com . (B) Quantitative comparison of protein abundance in the secretome versus the total proteome of MCF10A cells. Proteins with secretion changes reflecting a similar change in total expression (“explained by proteome”) (q value ≤ 0.05) are shown in green. Proteins with increased or decreased secretion independent of total proteome changes are highlighted in red and blue, respectively. Gray (not affected) represents unchanged proteins. (C) Analysis of protein abundance changes in the secretome of YIPF5 KO versus control MCF10A cells (WT). Proteins significantly upregulated (red) or downregulated (blue) in the YIPF5 KO secretome are shown. Proteins with secretion changes explained by total proteome abundance are shown in green, while unaffected proteins are in gray. The vertical axis represents –log10(q value), and the horizontal axis represents log2(fold change). (D and E) Heatmap depicting proteins showing increased secretion in the YIPF5 KO secretome with a gene ontology (GO) annotation for the ER (GO: 0007029) and the Golgi apparatus (GO: 0005794) (D) or that contain a KDEL-ER retrieval sequence (E). (F) MCF10A WT and YIPF5 KO cells stably expressing the inducible ER-Ca 2+ sensor GCampER were analyzed using live fluorescence microscopy at identical settings. (G) Mean ± SD fluorescence intensity of GCampER was quantified in MCF10A WT and YIPF5 KO cells in 72 wells per cell line from n = 3 independent experiments. Unpaired t test, two-tailed p < 0.0001. (H) HeLa cells stably expressing shScramble or shRNAs against YIPF5 were transiently transfected with CNPY3-mCherry, supernatants were collected and cell lysates prepared after 24 h followed by western blot analysis using a CNPY3 antibody. A representative blot of n = 3 independent experiments is shown. For full size blot see C. (I) Quantification of CNPY3-secretion from (H). Displayed are arbitrary units normalized to the values of shScramble-expressing cells from n = 3 independent experiments. One-way ANOVA with Tukey’s multiple comparisons test, with ∗ indicating p < 0.05. (J) Analysis of protein abundance changes in the secretome of YIPF5 KO re-expressing YIPF5 I98S mutant versus control MCF10A cells (WT). Proteins significantly upregulated (red) or downregulated (blue) in the YIPF5 I98S secretome are shown ( n = 4). Proteins with secretion changes explained by total proteome abundance are denoted in green, while unaffected proteins are in gray. (K) Venn Diagram of protein abundance changes in YIPF5-KO cells and YIPF5-KO re-expressing YIPF5 I98S cells. Proteins with decreased (blue) or increased (red) secretome abundance are shown. See also and , , , , and .

Article Snippet: CNPY3 rabbit polyclonal , Proteintech , Cat# 15215-1-AP; RRID: AB_11182172.

Techniques: Metabolic Labelling, Labeling, Purification, Mass Spectrometry, Comparison, Quantitative Proteomics, Expressing, Control, Sequencing, Stable Transfection, Fluorescence, Microscopy, Two Tailed Test, Transfection, Western Blot, Mutagenesis

Journal: Frontiers in Pharmacology

Article Title: Liraglutide Increases VEGF Expression via CNPY2-PERK Pathway Induced by Hypoxia/Reoxygenation Injury

doi: 10.3389/fphar.2019.00789

Figure Lengend Snippet: Effect of Lir on CNPY2 after H/R injury in HUVECs. (A) Protein secretion of CNPY2. (B) mRNA expression level of CNPY2 from each group. (C) Protein expression level of CNPY2 from each group. (D) mRNA expression level of CNPY2. (E) Protein expression level of CNPY2. Results were normalized to the percentage of calnexin expression. Data are mean ± SE. *P < 0.05, **P < 0.01 compared to Sham group; # P < 0.05, ## P < 0.01 compared to H/R group.

Article Snippet: The human CNPY2 enzyme-linked immunosorbent assay (ELISA) kit was purchased from Proteintech Group (Chicago, IL, USA).

Techniques: Expressing

Journal: Frontiers in Pharmacology

Article Title: Liraglutide Increases VEGF Expression via CNPY2-PERK Pathway Induced by Hypoxia/Reoxygenation Injury

doi: 10.3389/fphar.2019.00789

Figure Lengend Snippet: Effect of Lir on the CNPY2-PERK pathway induced by H/R injury in HUVECs. (A) mRNA expression levels of the CNPY2-PERK pathway proteins were determined by RT-PCR. (B) Protein expression levels of the CNPY2-PERK pathway proteins were determined by Western blot. mRNA expression levels of the CNPY2-PERK pathway proteins ATF4 (C) , HIF1α (D) , and VEGF (E) . Protein expression levels of CNPY2-PERK pathway proteins ATF4 (F) , HIF1α (G) , and VEGF (H) . Expression of β-actin was used for the loading control. Results were normalized to the percentage of β-actin expression. Data are mean ± SE. *P < 0.05, **P < 0.01 compared to Sham group; # P < 0.05, ## P < 0.01 compared to H/R group; $ P < 0.05, $$ P < 0.01 compared to H/R+Lir group.

Article Snippet: The human CNPY2 enzyme-linked immunosorbent assay (ELISA) kit was purchased from Proteintech Group (Chicago, IL, USA).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Control