Journal: Scientific Reports

Article Title: B7-H3 and GD2 overexpression as immunotherapeutic targets in retinoblastoma

doi: 10.1038/s41598-025-34331-6

Figure Lengend Snippet: Surface antigen and immune checkpoint molecule expression on two human retinoblastoma (RB) cell lines, Y79 and WERI-Rb-1, and normal retinal pigmented epithelial cell line (ARPE-19). ( a ) GD2 surface expression on Y79, WERI-Rb-1, and ARPE-19 cell lines. ( b ) CD171 surface expression on Y79, WERI-Rb-1, and ARPE-19 cell lines. ( c ) B7-H3 surface expression on Y79, WERI-Rb-1, and ARPE-19 cell lines. ( d ) PD-L1 surface expression on Y79, WERI-Rb-1, and ARPE-19 cell lines.

Article Snippet: The Y79 (ATCC Cat# HTB-18, RRID: CVCL_1893), and WERI-Rb-1 (ATCC Cat#HTB-169, RRID: CVCL_1792), are human retinoblastoma cell lines.

Techniques: Expressing

Journal: Cell Death & Disease

Article Title: Loss of ELF2 drives topotecan resistance in retinoblastoma revealed by genome-wide CRISPR-Cas9 screening

doi: 10.1038/s41419-025-08335-z

Figure Lengend Snippet: A The schematic diagram illustrates the workflow of genome-wide CRISPR-Cas9 knockout library screening (CRISPR: Clustered Regularly Interspaced Short Palindromic Repeats). B The scatter plot depicts the results for topotecan positively selected hits in the CRISPR-Cas9 screening, with the top 20 hits shown in red. C KEGG analysis of the top 50 topotecan positively selected hits identified through genome-wide CRISPR-Cas9 knockout screening. D Relative cell viability of WERI-Rb1 and Y79 cells following treatment with topotecan or vehicle for 96 hours ( n = 3). E , F ELF2 protein expression in WERI-Rb1 and Y79 cells under topotecan treatment ( n = 3). Data are presented as means ± SD. Statistical analysis was performed using two-tailed Student’s t test ( D ) or one-way ANOVA and Tukey’s multiple comparison test ( F ); ****p < 0.0001, ***p < 0.001, **p < 0.01, *p < 0.05.

Article Snippet: The human WERI-Rb1 and Y79 cell lines were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Genome Wide, CRISPR, Knock-Out, Library Screening, Expressing, Two Tailed Test, Comparison

Journal: Cell Death & Disease

Article Title: Loss of ELF2 drives topotecan resistance in retinoblastoma revealed by genome-wide CRISPR-Cas9 screening

doi: 10.1038/s41419-025-08335-z

Figure Lengend Snippet: A A reduction of ELF2 protein expression in ELF2 knockout (KO; sgELF2) cells. B Western blot analysis of total caspase-3 and cleaved caspase-3 in topotecan (TPT)-treated ELF2 KO (sgELF2) and non-targeting control (NTC) cells. C , D Quantitative analysis of apoptotic cells by TUNEL assay in topotecan-treated ELF2 KO (sgELF2) and NTC cells ( n = 3; scale bar: 50 µm). The relative TUNEL-positive rate was normalized to the mean value of the NT + NaCl group (set as 1.0). E Representative western blot image of ELF2 and Caspase-3 proteins in the ELF2-overexpressing (pCMV3-ELF2-t3) and control (pCMV3-untagged) WERI-Rb1 cells. F , G Quantitative analysis of ELF2 protein, cleaved caspase-3 proteins and total caspase-3 ( n = 3). Bar groups in E–G represent: 1 = pCMV3-untagged, 2 = pCMV3-untagged + TPT, 3 = pCMV3-ELF2-t3, 4 = pCMV3-ELF2-t3 + TPT. H Relative cell viability of control and ELF2 overexpressing cells following treatment with topotecan for 72 h ( n = 3). In H , the bars represent fold-change comparisons: 2/1 = (pCMV3-untagged + TPT)/(pCMV3-untagged); 4/3 = (pCMV3-ELF2-t3 + TPT)/(pCMV3-ELF2-t3). I and J Quantitative analysis of apoptotic cells by TUNEL assay in topotecan-treated ELF2 overexpressing and control cells ( n = 3; scale bar: 50 µm). Data are presented as means ± SD. Statistical analysis was performed using two-tailed Student’s t test ( A , H ) or one-way ANOVA and Tukey’s multiple comparison test ( B , D , F , G and J ); **** p < 0.0001, *** p < 0.001, ** p < 0.01, * p < 0.05.

Article Snippet: The human WERI-Rb1 and Y79 cell lines were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Expressing, Knock-Out, Western Blot, Control, TUNEL Assay, Two Tailed Test, Comparison

Journal: Cell Death & Disease

Article Title: Loss of ELF2 drives topotecan resistance in retinoblastoma revealed by genome-wide CRISPR-Cas9 screening

doi: 10.1038/s41419-025-08335-z

Figure Lengend Snippet: A Workflow of in vivo mouse xenotransplantation study. Seven days after the transplantation of non-targeting control (NTC) or ELF2 knockout (sgELF2) WERI-Rb1 cells (denoted as day 0), mice were subjected to an intraperitoneal injection of saline or 0.1 mg/kg topotecan (TPT) once daily, Monday through Friday, for 3 consecutive weeks (total 15 doses). B Following the initiation of dosing, each mouse was monitored with callipers, and tumor volumes were plotted graphically ( n = 6). C The macroscopic appearance of xenotransplanted tumors 21 days following topotecan treatment. D and E Representative western blot image and quantitative analysis of ELF2 and Caspase-3 proteins in topotecan-treated ELF2 KO (sgELF2) and NTC tumor tissues ( n = 6). F , G Representative image and quantitative analysis of apoptotic cells by TUNEL assay in topotecan-treated ELF2 KO (sgELF2) and NTC tumor tissues ( n = 6; scale bar: 50 µm). Data are presented as means ± SD. Statistical analysis was performed using one-way ANOVA and Tukey’s multiple comparison test ( B , E and G ); **** p < 0.0001, ** p < 0.01, * p < 0.05.

Article Snippet: The human WERI-Rb1 and Y79 cell lines were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: In Vivo, Transplantation Assay, Control, Knock-Out, Injection, Saline, Western Blot, TUNEL Assay, Comparison

Journal: Cell Death & Disease

Article Title: Loss of ELF2 drives topotecan resistance in retinoblastoma revealed by genome-wide CRISPR-Cas9 screening

doi: 10.1038/s41419-025-08335-z

Figure Lengend Snippet: A Principal component analysis (PCA) of RNA-seq data from the experimental groups (non-targeting control (NTC), ELF2 knockout (KO; sgELF2), topotecan(TPT)-treated NTC, topotecan-treated ELF2 KO WERI-Rb1 cells; n = 3). B Volcano plots illustrate the genes that were significantly altered in both NTC and ELF2 KO (sgELF2) WERI-Rb1 cells upon stimulation with topotecan. Red and blue dots indicate genes exhibiting a log2|fold change | > 1 and FDR < 0.05. C Venn diagram of differentially expressed genes (DEGs) between the two data sets. D Gene Ontology and E KEGG enrichment analysis of the identified genes from ( C ).

Article Snippet: The human WERI-Rb1 and Y79 cell lines were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: RNA Sequencing, Control, Knock-Out

Journal: Cell Death & Disease

Article Title: Loss of ELF2 drives topotecan resistance in retinoblastoma revealed by genome-wide CRISPR-Cas9 screening

doi: 10.1038/s41419-025-08335-z

Figure Lengend Snippet: A Heatmap illustrating the expression of 17 genes involved in the metabolic pathways identified in Fig. for each sample across various groups. B The mRNA levels of MT-CYB in WERI-Rb1 cells across different groups. C The mRNA levels of MT-CYB in Y79 cells across different groups. D Heatmap illustrating the expression of 12 mtDNA-encoded genes across groups. E GSEA enrichment plots of OXIDATIVE_PHOSPHORYLATION pathways comparing sgELF2 + topotecan(TPT) vs. NTC + TPT. F , G Representative Western blot and quantitative analysis of MT-CYB expression in control (NTC + pCMV3-untagged), ELF2 knockout (sgELF2 + pCMV3-untagged), and ELF2 rescue (sgELF2 + pCMV3-ELF2-t3) cells. H , I Quantitative analysis of mitochondrial DNA copy number and cellular ATP levels in WERI-Rb1 cells across various groups. J The relative contribution of mitochondrial and glycolytic ATP across different groups. Data are presented as means ± SD. Statistical analysis was performed using one-way ANOVA and Tukey’s multiple comparison test ( B , C , G , H , I and J ); **** p < 0.0001, *** p < 0.001, ** p < 0.01, * p < 0.05.

Article Snippet: The human WERI-Rb1 and Y79 cell lines were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Expressing, Phospho-proteomics, Western Blot, Control, Knock-Out, Comparison