Journal: Oncology Letters

Article Title: Exploring multiple biomarkers and constructing ferroptosis-associated competing endogenous RNA networks as dual targets in retinoblastoma

doi: 10.3892/ol.2026.15582

Figure Lengend Snippet: ceRNA networks, drug-gene interactions and PCR validation. (A) The ceRNA-regulating networks illustrate protein-coding genes (red circles), miRNAs (blue diamonds) and lncRNAs (green hexagons), with black lines indicating interactions among lncRNA, miRNA and mRNA, where ceRNA refers to ceRNAs. (B) Validation through reverse transcription-quantitative PCR analysis confirms that CDKN2A and IDH2 expression levels are significantly higher in Y79 cells compared to ARPE-19 cells. Data were normalized to GAPDH expression using the 2 −ΔΔCq method, with each experiment performed in triplicate. ***P<0.001 and ****P<0.0001. (C) The drug-gene interaction analysis shows the linkage map of two hub genes, CDKN2A and IDH2 , along with their potential target drugs, including bisphenol A (code: C006780), sodium arsenite (code: C017947), docetaxel (code: D000077143) and hexabromocyclododecane (code: C089796). ceRNA, competing endogenous RNAs; miRNA, microRNA; lncRNA, long non-coding RNA; RB, retinoblastoma.

Article Snippet: The human retinal pigment epithelium cell line ARPE-19 and the human RB cell line Y79 were sourced from the American Type Culture Collection.

Techniques: Biomarker Discovery, Reverse Transcription, Real-time Polymerase Chain Reaction, Expressing

Journal: The Journal of Biological Chemistry

Article Title: Age-dependent induction of ER stress in retinal pigment epithelium impairs phagocytosis via ADAM17-dependent MERTK shedding

doi: 10.1016/j.jbc.2026.111397

Figure Lengend Snippet: Pharmacological induction of ER stress attenuates phagocytic activity in cultured RPE cells. A , schematic diagram of the phagocytosis assay using fluorescein isothiocyanate (FITC)- and pHrodo succinimidyl ester (pHrodo)-conjugated photoreceptor outer segments (POS). B , a representative image of engulfed FITC-POS ( green ) and Hoechst 33,342 ( blue ) with plasma membrane staining 6 h after FITC-POS treatment. The plasma membrane ( gray ) was visualized by PlasMem Bright Red. Scale bar = 10 μm. C , a representative image of pHrodo signal ( yellow ), LAMP1 (magenta) at 24 h after pHrodo-POS treatment. Scale bar = 10 μm. D–F , Tunicamycin (Tm)-induced short-term ER stress reduces phagocytic activity in ARPE-19 and human primary RPE (hRPE) cells. D , experimental timeline for the assays shown in ( E ) and ( F ). E , quantification of fluorescence intensity for FITC-POS and pHrodo-POS in ARPE-19. Data are presented as mean ± SEM (n = 6). Each point represents one independent sample prepared from separate wells. ### p < 0.001 vs. control (Cont) group (Dunnett’s test). F , quantitative data of fluorescence intensity for pHrodo-POS in hRPE cells. Data are presented as mean ± SEM (n = 6). Each point represents one independent sample prepared from separate wells. ## p < 0.01, ### p < 0.001 vs. Cont group (Dunnett’s test). G , quantification of phagocytized pHrodo-POS after co-treatment with thapsigargin (Tg). Data are presented as mean ± SEM (n = 6). Each point represents one independent sample prepared from separate wells. # p < 0.05 vs. Cont group (Student's t test). H , cell death rate following Tm or Tg treatment for 6 h. Data are presented as mean ± SEM (n = 6). Each point represents one independent sample prepared from separate wells. N.S. > 0.05 vs. Cont group (Dunnett’s test). I – K , long-term ER stress reduces phagocytic activity in ARPE -19 and hRPE. I , experimental timelines for assays shown in ( J ) and ( K ). Quantitative data of fluorescence intensity of pHrodo-POS in ARPE-19 ( J ) and hRPE ( K ). Data are presented as mean ± SEM (n = 6). Each point represents one independent sample prepared from separate wells. # p < 0.05, ## p < 0.01, ### p < 0.001 vs. Cont group (Dunnett’s test).

Article Snippet: The human-derived RPE cell line, ARPE-19, was purchased from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Activity Assay, Cell Culture, Phagocytosis Assay, Clinical Proteomics, Membrane, Staining, Fluorescence, Control

Journal: The Journal of Biological Chemistry

Article Title: Age-dependent induction of ER stress in retinal pigment epithelium impairs phagocytosis via ADAM17-dependent MERTK shedding

doi: 10.1016/j.jbc.2026.111397

Figure Lengend Snippet: ER stress induces lysosomal and mitochondrial dysfunction in ARPE-19 cells. A – F , effect of Tm on lysosomal activity in ARPE-19 cells. Representative images of LysoTracker Red DND-99 staining at 3 h ( A ), 6 h ( C ), and 54 h ( E ) post-Tm treatment. Scale bar = 200 μm. Quantification of LysoTracker fluorescence intensity at 3 h ( B ), 6 h ( D ), and 54 h ( F ) post-Tm treatment. Data are presented as mean ± SEM (n = 6). Each point represents one independent well. ## p < 0.01, ### p < 0.001 vs. Cont group (Dunnett’s T3 test). ( G ) Representative kinetic graph of mitochondrial stress test in ARPE-19 cells, three or 6 h post-Tm treatment. Quantification of mitochondrial basal respiration ( H ) and maximum respiration ( I ) in ARPE-19. Oligomycin (Oligo), ATP synthase inhibitor; FCCP, mitochondrial respiration uncoupler to reveal maximal respiratory capacity; rotenone/antimycin A (Rot/A), complexes I and III inhibitor to define non-mitochondrial respiration. Data are presented as mean ± SEM (n = 4). Each point represents one independent well. # p < 0.05, ## p < 0.01 vs. Cont group (Dunnett’s test).

Article Snippet: The human-derived RPE cell line, ARPE-19, was purchased from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Activity Assay, Staining, Fluorescence

Journal: The Journal of Biological Chemistry

Article Title: Age-dependent induction of ER stress in retinal pigment epithelium impairs phagocytosis via ADAM17-dependent MERTK shedding

doi: 10.1016/j.jbc.2026.111397

Figure Lengend Snippet: Effect of ER stress reduction on phagocytic impairment induced by Tm. A – C , effect of sodium 4-phenylbutyrate (4-PBA) on Tm-induced phagocytic impairment. A , experimental protocol for the assay shown in ( B ). Quantitative data of fluorescence intensity for pHrodo-POS ( B ) and LysoTracker ( C ). Data are presented as mean ± SEM (n = 6). Each point represents one independent well. # p < 0.05, ### p < 0.001 vs Cont group; ∗ p < 0.05 vs. Tm single-treated group (Tukey's test). D–F , induction of activating transcription factor 6 nuclear form (ATF6n), XBP1s, and chaperone proteins 2 days post-plasmid transfection in ARPE-19. D , representative immunoblots of ATF6n and β-actin. E , quantitative data on ATF6n expression normalized to β-actin. Data are presented as mean ± SEM (n = 3). Each point represents one independent sample prepared from separate wells. # p < 0.05 vs. Mock plasmid-transfected group (Student’s t test). F , gene expression of XBP1s , glucose-regulated protein 94 ( GRP94 ), and GRP78 normalized to β-actin 2 days after plasmid transfection. Data are presented as mean ± SEM (n = 5). Each point represents one independent sample prepared from separate wells. # p < 0.05, ## p < 0.01, ### p < 0.001 vs. Mock plasmid-transfected group (Tukey’s test). G and H , effect of chaperone-inducible plasmids on Tm-induced phagocytic impairment in ARPE-19. G , experimental protocol for the assay shown in ( H ). H , quantitative data of fluorescence intensity for pHrodo-POS in plasmid-transfected groups. Data are presented as mean ± SEM (n = 6). Each point represents one independent well. # p < 0.05, ## p < 0.01 vs. Mock plasmid single-treated group; ∗ p < 0.05, ∗∗ p < 0.05 vs. Mock plasmid and Tm co-treated group (Tukey’s test).

Article Snippet: The human-derived RPE cell line, ARPE-19, was purchased from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Fluorescence, Plasmid Preparation, Transfection, Western Blot, Expressing, Gene Expression

Journal: The Journal of Biological Chemistry

Article Title: Age-dependent induction of ER stress in retinal pigment epithelium impairs phagocytosis via ADAM17-dependent MERTK shedding

doi: 10.1016/j.jbc.2026.111397

Figure Lengend Snippet: ER stress induces Mer tyrosine kinase receptor (MERTK) shedding and downregulation of phagocytosis-related signaling. A , schematic diagram of POS internalized step mediated by integrin αvβ5, MERTK and focal adhesion kinase (FAK). B and C , expression changes of ER stress-related and phagocytosis-related proteins after short-term Tm (10 μg/ml, 6 h) treatment. B , representative immunoblots of p-eIF2α, eIF2α, ATF4, GRP78, phosphorylated MERTK (p-MERTK), FAK, phosphorylated FAK (p-FAK), MERTK (Full length) detected by antibody against N-terminal (N-ter), and β-actin. C , quantitative data for the factors above normalized to β-actin or corresponded non-phosphorylated protein. Data are presented as mean ± SEM (n = 6). Each point represents one independent sample prepared from separate wells. # p < 0.05, ## p < 0.01, ### p < 0.001 vs. Cont group (Welch's t test). D , time-dependent change in GRP78 and MERTK mRNA expression normalized to β-actin after Tm treatment. Data are presented as mean ± SEM (n = 4). Each point represents one independent sample prepared from separate wells. # p < 0.05, ### p < 0.001 vs. Cont group (Dunnett’s T3 test). E and F , changes in MERTK expression in lysate from hRPE after Tm treatment (10 μg/ml, 6 h), distinguished by glycosylation status based on molecular weight. E , representative immunoblots of MERTK (Extracellular domain) detected by antibody against N-terminal (N-ter). The band observed at approximately 170 kDa corresponds to the glycosylated full-length form of MERTK, whereas the band at approximately 110 kDa represents the un-glycosylated form of MERTK, as predicted from its amino acid sequence. F , quantitative analysis of the expression levels of glycosylated full-length MERTK and un-glycosylated MERTK, normalized to β-actin, as well as their ratio. Data are presented as mean ± SEM (n = 6). Each point represents one independent sample prepared from separate wells. ## p < 0.01, ### p < 0.001 vs. Cont group (Student’s t test). G , quantification by enzyme-linked immunosorbent assay of the extracellular domain of MERTK in the culture medium of Tm-treated ARPE-19 and hRPE. Data are presented as mean ± SEM (n = 6). Each point represents one independent sample prepared from separate wells. ## p < 0.05 vs. Cont group (Welch’s t test). H and I , the expression of released MERTK having diverse molecular weight in culture medium of Tm (10 μg/ml, 6 h)-treated hRPE cells detected by antibody against extracellular domain (N-ter) of MERTK. H , representative immunoblots of MERTK (Extracellular domain). The bands correspond as follow: approximately 170 kDa, full-length MERTK; approximately 120 kDa, soluble MERTK generated by shedding; approximately 65 kDa, un-glycosylated soluble MERTK; approximately 50 kDa is an unknown band, observed only in the culture medium sample compared to the lysate sample ( E ). F , quantitative analysis of the expression levels of full-length MERTK and soluble MERTK in culture medium Tm-treated hRPE. Data are presented as mean ± SEM (n = 4). Each data point represents one independent sample prepared by pooling conditioned medium from two separate wells. # p < 0.05, ## p < 0.01 vs. Cont group (Student’s t test).

Article Snippet: The human-derived RPE cell line, ARPE-19, was purchased from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Expressing, Western Blot, Glycoproteomics, Molecular Weight, Sequencing, Enzyme-linked Immunosorbent Assay, Generated

Journal: The Journal of Biological Chemistry

Article Title: Age-dependent induction of ER stress in retinal pigment epithelium impairs phagocytosis via ADAM17-dependent MERTK shedding

doi: 10.1016/j.jbc.2026.111397

Figure Lengend Snippet: Maturation of ADAM17 mediated by Ca 2+ release from inositol - 1,4,5-trisphosphate receptors contributes to MERTK shedding and dysfunction of POS uptake. A , time-dependent change of the mature form of ADAM17 (matADAM17) in ARPE-19 cells after Tm treatment at 10 μg/ml. Data are presented as mean ± SEM (n = 4). Each point represents one independent sample prepared from separate wells. # p < 0.05 vs. Cont group (Welch's t test). B , expression level of matADAM17 in ARPE-19 after Tm treatment at 1 μg/ml for 54 h. Data are presented as mean ± SEM (n = 6). Each point represents one independent sample prepared from separate wells. ### p < 0.001 vs. Cont group (Welch’s t test). C , localization of ADAM17 after Tm treatment at 10 μg/ml for 3 h. Representative images of ADAM17 ( yellow ), Golgin-97 ( magenta ), and Hoechst 33,342 ( blue ). Scale bar = 10 μm. Quantitative data of fluorescence intensity of ADAM17 colocalized with Golgin-97 after 1 and 3 h after Tm treatment. Data are presented as mean ± SEM (Cont; n = 97 cells, Tm 1 h; n = 103 cells, Tm 3 h; n = 104 cells). ### p < 0.001 vs. Cont group (Dunnett's T3 test). D – F , expression level of matADAM17 after Tm treatment at 10 μg/ml in the presence of decanoyl-Arg-Val-Lys-Arg-chloromethylketone (CMK) (100 μM, D), 1,2-bis(o-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid, tetraacetoxymethyl ester (BAPTA-AM) (100 μM, E ), or 2-aminoethoxydiphenyl borate (2-APB) (100 μM, F ). Data are presented as mean ± SEM (n = 6). Each point represents one independent sample prepared from separate wells. ## p < 0.01, ### p < 0.001 vs. Cont group; ∗∗ p < 0.01, ∗∗∗ p < 0.001 vs. Tm-only treated group (Games–Howell test). G , schematic diagram of ER stress-induced ADAM17 maturation and MERTK shedding. H and I , effect of ADAM17 small interfering RNA (siRNA) treatment on Tm-induced MERTK downregulation in ARPE-19. H , representative immunoblots of MERTK (extracellular domain), ADAM17, and β-actin. I , quantitative data for MERTK (extracellular domain). Data are presented as mean ± SEM (n = 6). Each point represents one independent sample prepared from separate wells. ## p < 0.01 vs. control siRNA (siCont) single-treated group; ∗ p < 0.05 vs. siCont and Tm co-treated group (Student's t test). J and K , effect of ADAM17 siRNA on Tm-induced dysfunction of POS uptake in ARPE-19. J , schematic protocol of the POS uptake assay and ( K ) quantitative data of fluorescence intensity of FITC-POS internalized in RPE cells. Data are presented as mean ± SEM (n = 8). Each point represents one independent well. # p < 0.05 vs. siCont single-treated group; †† p < 0.01 vs. siCont and chloroquine co-treated group; ∗ p < 0.05 vs. siCont, chloroquine, and Tm co-treated group (Kruskal–Wallis test followed by post hoc Bonferroni test).

Article Snippet: The human-derived RPE cell line, ARPE-19, was purchased from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Expressing, Fluorescence, Small Interfering RNA, Western Blot, Control

Journal: The Journal of Biological Chemistry

Article Title: Age-dependent induction of ER stress in retinal pigment epithelium impairs phagocytosis via ADAM17-dependent MERTK shedding

doi: 10.1016/j.jbc.2026.111397

Figure Lengend Snippet: Pharmacological induction of ER stress attenuates phagocytic activity in cultured RPE cells. A , schematic diagram of the phagocytosis assay using fluorescein isothiocyanate (FITC)- and pHrodo succinimidyl ester (pHrodo)-conjugated photoreceptor outer segments (POS). B , a representative image of engulfed FITC-POS ( green ) and Hoechst 33,342 ( blue ) with plasma membrane staining 6 h after FITC-POS treatment. The plasma membrane ( gray ) was visualized by PlasMem Bright Red. Scale bar = 10 μm. C , a representative image of pHrodo signal ( yellow ), LAMP1 (magenta) at 24 h after pHrodo-POS treatment. Scale bar = 10 μm. D–F , Tunicamycin (Tm)-induced short-term ER stress reduces phagocytic activity in ARPE-19 and human primary RPE (hRPE) cells. D , experimental timeline for the assays shown in ( E ) and ( F ). E , quantification of fluorescence intensity for FITC-POS and pHrodo-POS in ARPE-19. Data are presented as mean ± SEM (n = 6). Each point represents one independent sample prepared from separate wells. ### p < 0.001 vs. control (Cont) group (Dunnett’s test). F , quantitative data of fluorescence intensity for pHrodo-POS in hRPE cells. Data are presented as mean ± SEM (n = 6). Each point represents one independent sample prepared from separate wells. ## p < 0.01, ### p < 0.001 vs. Cont group (Dunnett’s test). G , quantification of phagocytized pHrodo-POS after co-treatment with thapsigargin (Tg). Data are presented as mean ± SEM (n = 6). Each point represents one independent sample prepared from separate wells. # p < 0.05 vs. Cont group (Student's t test). H , cell death rate following Tm or Tg treatment for 6 h. Data are presented as mean ± SEM (n = 6). Each point represents one independent sample prepared from separate wells. N.S. > 0.05 vs. Cont group (Dunnett’s test). I – K , long-term ER stress reduces phagocytic activity in ARPE -19 and hRPE. I , experimental timelines for assays shown in ( J ) and ( K ). Quantitative data of fluorescence intensity of pHrodo-POS in ARPE-19 ( J ) and hRPE ( K ). Data are presented as mean ± SEM (n = 6). Each point represents one independent sample prepared from separate wells. # p < 0.05, ## p < 0.01, ### p < 0.001 vs. Cont group (Dunnett’s test).

Article Snippet: The human-derived RPE cell line, ARPE-19, was purchased from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Activity Assay, Cell Culture, Phagocytosis Assay, Clinical Proteomics, Membrane, Staining, Fluorescence, Control

Journal: The Journal of Biological Chemistry

Article Title: Age-dependent induction of ER stress in retinal pigment epithelium impairs phagocytosis via ADAM17-dependent MERTK shedding

doi: 10.1016/j.jbc.2026.111397

Figure Lengend Snippet: Maturation of ADAM17 mediated by Ca 2+ release from inositol - 1,4,5-trisphosphate receptors contributes to MERTK shedding and dysfunction of POS uptake. A , time-dependent change of the mature form of ADAM17 (matADAM17) in ARPE-19 cells after Tm treatment at 10 μg/ml. Data are presented as mean ± SEM (n = 4). Each point represents one independent sample prepared from separate wells. # p < 0.05 vs. Cont group (Welch's t test). B , expression level of matADAM17 in ARPE-19 after Tm treatment at 1 μg/ml for 54 h. Data are presented as mean ± SEM (n = 6). Each point represents one independent sample prepared from separate wells. ### p < 0.001 vs. Cont group (Welch’s t test). C , localization of ADAM17 after Tm treatment at 10 μg/ml for 3 h. Representative images of ADAM17 ( yellow ), Golgin-97 ( magenta ), and Hoechst 33,342 ( blue ). Scale bar = 10 μm. Quantitative data of fluorescence intensity of ADAM17 colocalized with Golgin-97 after 1 and 3 h after Tm treatment. Data are presented as mean ± SEM (Cont; n = 97 cells, Tm 1 h; n = 103 cells, Tm 3 h; n = 104 cells). ### p < 0.001 vs. Cont group (Dunnett's T3 test). D – F , expression level of matADAM17 after Tm treatment at 10 μg/ml in the presence of decanoyl-Arg-Val-Lys-Arg-chloromethylketone (CMK) (100 μM, D), 1,2-bis(o-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid, tetraacetoxymethyl ester (BAPTA-AM) (100 μM, E ), or 2-aminoethoxydiphenyl borate (2-APB) (100 μM, F ). Data are presented as mean ± SEM (n = 6). Each point represents one independent sample prepared from separate wells. ## p < 0.01, ### p < 0.001 vs. Cont group; ∗∗ p < 0.01, ∗∗∗ p < 0.001 vs. Tm-only treated group (Games–Howell test). G , schematic diagram of ER stress-induced ADAM17 maturation and MERTK shedding. H and I , effect of ADAM17 small interfering RNA (siRNA) treatment on Tm-induced MERTK downregulation in ARPE-19. H , representative immunoblots of MERTK (extracellular domain), ADAM17, and β-actin. I , quantitative data for MERTK (extracellular domain). Data are presented as mean ± SEM (n = 6). Each point represents one independent sample prepared from separate wells. ## p < 0.01 vs. control siRNA (siCont) single-treated group; ∗ p < 0.05 vs. siCont and Tm co-treated group (Student's t test). J and K , effect of ADAM17 siRNA on Tm-induced dysfunction of POS uptake in ARPE-19. J , schematic protocol of the POS uptake assay and ( K ) quantitative data of fluorescence intensity of FITC-POS internalized in RPE cells. Data are presented as mean ± SEM (n = 8). Each point represents one independent well. # p < 0.05 vs. siCont single-treated group; †† p < 0.01 vs. siCont and chloroquine co-treated group; ∗ p < 0.05 vs. siCont, chloroquine, and Tm co-treated group (Kruskal–Wallis test followed by post hoc Bonferroni test).

Article Snippet: The human-derived RPE cell line, ARPE-19, was purchased from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Expressing, Fluorescence, Small Interfering RNA, Western Blot, Control

Journal: Archives of Gynecology and Obstetrics

Article Title: Possible role of hyperimmunoglobulin in reducing the risk of maternal–fetal transmission of cytomegalovirus in the valacyclovir era: a case series

doi: 10.1007/s00404-026-08432-0

Figure Lengend Snippet: CMV-specific antibody levels and activity in case 3. Anti-CMV IgG (circles) and IgM (squares) levels at different days after the first detection of CMV-DNA are shown in panel A. The serum neutralization titer against the infection of epithelial (ARPE-19, circles) and fibroblast (MRC-5, squares) cells and the antibody-dependent cell-cytotoxicity (ADCC) against the infection of ARPE-19 cells are shown in panels B and C, respectively. Vertical dotted lines represent the days of the HIG administration. Red points illustrate tests performed right after HIG administration

Article Snippet: For the neutralization assay, we used VR1814 for epithelial cells (ARPE-19, ATCC) and AD169 (ATCC, Manassas, VA, USA), a reference laboratory-adapted strain, for fibroblast cells (HELF, isolated in-house).

Techniques: Activity Assay, Neutralization, Infection