Journal: bioRxiv

Article Title: A Novel Therapeutic Approach: Notch Inhibition Enhances Radiotherapy and Checkpoint Blockade Therapy via Reprogramming of the Tumor Microenvironment

doi: 10.64898/2025.12.30.696685

Figure Lengend Snippet: (A). Tumor cells were implanted into the flank (1 x 10 6 9464D) or mammary fat pad (1 x 10 5 EO771) of C57BL/6 mice, enrolled when tumors reached 0.15-0.2mm 3 . Mice were treated as per the schema and sacrificed when tumors reached a size 1.5cm 3 . (B) The Kaplan-Meier survival curve of mice and tumor volume growth curve of 9464D tumor. Median Survival: Control 12d; IgG 10d; GSI 10d; aPD-1 10d; aPD-1+ GSI 10d; RT 37d; RT+ GSI 50d; RT+aPD-1 46d; RT+aPD-1+AL101 87d. Curve comparison by Log-rank (Mantel-Cox) test. P<0.0001. (C) The Kaplan-Meier survival curve of mice and tumor volume growth curve of EO771 tumor Median Survival: Control: 14d; IgG 14.5d; GSI 18.5d; aPD-1 15d; GSI+aPD-1 10d; RT 35d; RT+ GSI 41d; RT+aPD-1 43d; RT+aPD-1+ GSI 98.5d. Curve comparison by Log-rank (Mantel-Cox) test. P<0.0001. (D) Lungs were collected from EO771 tumor–bearing mice (n = 4–7 per group) treated as described above and sacrificed either at day 10 or at the survival endpoint. Tumor metastases were quantified using HALO image analysis, calculated as metastatic area divided by total lung area. Bars represent mean ± SD. *P<0.05, using one-way ANOVA followed by Tukey post-hoc comparisons.

Article Snippet: EO771 cells (purchased from ATCC) and the 9464D cells (derived from a TH-MYCN transgenic neuroblastoma mouse, obtained as a gift from Dr.

Techniques: Control, Comparison

Journal: bioRxiv

Article Title: A Novel Therapeutic Approach: Notch Inhibition Enhances Radiotherapy and Checkpoint Blockade Therapy via Reprogramming of the Tumor Microenvironment

doi: 10.64898/2025.12.30.696685

Figure Lengend Snippet: (A) UMAPs showing M1-like and M2-like macrophage states across treatments in 9464D tumor scRNA-seq. (B) Proportion of M1- and M2-like macrophages by treatment group (C) Expression of classical M1 (Il1b, Tnf) and M2 (Mrc1, Mgl2) markers. (D) M-MDSC module scores increase following RT + aPD-1 + GSI (E) Changes in the proportion of M1 and M2 macrophages subpopulation and Myeloid-derived suppressor cells (MDSC) subpopulations by flow cytometry in 9464D tumor (F) Changes in the proportion of M1 and M2 macrophages subpopulation and MDSC subpopulations by flow cytometry in EO771 tumor. Bars represent the mean. Each dot represents one animal. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001 using one-way ANOVA followed by Tukey post-hoc comparisons.

Article Snippet: EO771 cells (purchased from ATCC) and the 9464D cells (derived from a TH-MYCN transgenic neuroblastoma mouse, obtained as a gift from Dr.

Techniques: Expressing, Derivative Assay, Flow Cytometry

Journal: bioRxiv

Article Title: A Novel Therapeutic Approach: Notch Inhibition Enhances Radiotherapy and Checkpoint Blockade Therapy via Reprogramming of the Tumor Microenvironment

doi: 10.64898/2025.12.30.696685

Figure Lengend Snippet: (A) Triple therapy enhances DC activation and antigen-presentation gene expression in 9464D tumor scRNA-seq. (B) Changes in the proportion of DC and CD103+DC proportion by flowcytometry in 9464D tumor. (C) Changes in the proportion of DC and CD103+DC proportion by flowcytometry in EO771 tumor. Bars represent mean, each dot represents one animal. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001 using one-way ANOVA followed by Tukey post-hoc comparisons.

Article Snippet: EO771 cells (purchased from ATCC) and the 9464D cells (derived from a TH-MYCN transgenic neuroblastoma mouse, obtained as a gift from Dr.

Techniques: Activation Assay, Immunopeptidomics, Gene Expression

Journal: bioRxiv

Article Title: A Novel Therapeutic Approach: Notch Inhibition Enhances Radiotherapy and Checkpoint Blockade Therapy via Reprogramming of the Tumor Microenvironment

doi: 10.64898/2025.12.30.696685

Figure Lengend Snippet: 9464D cells or EO771 were implanted into NCr nude mice and treated per the schema in . Mice were sacrificed when tumors reached a size >1.5 cm 3 . (A) The Kaplan-Meier survival curve of mice and tumor volume growth curve of athymic nude mice 9464D tumor. (B) The Kaplan-Meier survival curve of mice and tumor volume growth curve, as well as lung metastasis of athymic nude mice EO771 tumor. Comparison by Log-rank (Mantel-Cox) test and one-way ANOVA.

Article Snippet: EO771 cells (purchased from ATCC) and the 9464D cells (derived from a TH-MYCN transgenic neuroblastoma mouse, obtained as a gift from Dr.

Techniques: Comparison