niclosamide  (MedChemExpress)

Journal: bioRxiv

Article Title: Identifying the therapeutic potential of Niclosamide in overcoming IFN-gamma dependent cancer immune evasion in the Tumor Microenvironment

doi: 10.1101/2025.10.24.684401

Figure Lengend Snippet: PD-L1 expression on MC38 cell surface was significantly increased in the presence of IFN-γ analyzed by flow cytometry ( A ) and by immune fluorescent staining ( B - C ) at 24 hours. ( D ) When treated with siRNA of STAT1, the up-regulation of PD-L1 was significantly reduced, while siRNA of STAT3 has an opposite effect. ( E-F ) The small interfering RNA reaches a sufficient knock down of STAT1 and STAT3, as shown by real-time PCR assay. ( G-I ) The mRNA of STAT1 target genes such as CXCL10 and IL-15 shown similar trend of regulation by IFN-γ as PD-L1. ( J ) The mRNA of STAT3 target gene, MCL-1, shown totally different regulation under normal condition and under treatment of siRNAs. ( K-L) The expression of STAT1/STAT3 normal and phosphorylated proteins, and PD-L1 proteins in MC38 cell lines were measured by Western blotting with different dose of IFN-γ stimulation (K ) or after knocking down of STAT1 (siSTAT1) or STAT3 (siSTAT3) (L ). The number on the image indicates the relative abundance of PD-L1 protein (fold of control). The results are expressed as the mean ± SEM of triplicate measurements in each group. *p<0.05, **p<0.01, ***p<0.001.

Article Snippet: We evaluated small molecule STAT1 inhibitor Fludarabine (F-ara-A, NSC 118218) (MedChemExpress, Cat. No.: HY-B0069) and STAT3 inhibitor Niclosamide (BAY2353, MedChemExpress, Cat. No.: HY-B0497).

Techniques: Expressing, Flow Cytometry, Staining, Small Interfering RNA, Knockdown, Real-time Polymerase Chain Reaction, Western Blot, Control

Journal: bioRxiv

Article Title: Identifying the therapeutic potential of Niclosamide in overcoming IFN-gamma dependent cancer immune evasion in the Tumor Microenvironment

doi: 10.1101/2025.10.24.684401

Figure Lengend Snippet: ( A-C ) Ki67 expression in the MC38 tumor spheres was significantly reduced in the presence of IFN-γ analyzed by flow cytometry and immune fluorescence. ( D-E ) The tumor sphere forming unit induced in the MC38 cells were also reduced in the presence of IFN-γ (Arrowhead: tumor spheroid. Scale bar: 200μm). (F) When MC38 cells were pre-treated with siRNA of STAT1/STAT3, the SFU show different changes. (G) When MC38 cells were pre-treated with siRNA of STAT1/STAT3, the CD44+CD133+ population show different trend of regulation by IFN-γ. (H-K) The mRNA of cancer stem cell markers was measured by real-time PCR. (L) The expression of cancer stem cell marker proteins in cell nucleus was measured by Western blotting. The results are expressed as the mean ± SEM of triplicate measurements in each group. *p<0.05, **p<0.01, ***p<0.001.

Article Snippet: We evaluated small molecule STAT1 inhibitor Fludarabine (F-ara-A, NSC 118218) (MedChemExpress, Cat. No.: HY-B0069) and STAT3 inhibitor Niclosamide (BAY2353, MedChemExpress, Cat. No.: HY-B0497).

Techniques: Expressing, Flow Cytometry, Fluorescence, Real-time Polymerase Chain Reaction, Marker, Western Blot

Journal: bioRxiv

Article Title: Identifying the therapeutic potential of Niclosamide in overcoming IFN-gamma dependent cancer immune evasion in the Tumor Microenvironment

doi: 10.1101/2025.10.24.684401

Figure Lengend Snippet: (A) The expression of STAT1/STAT3 signaling pathway and PD-L1 proteins in MC38 cell lines with Fludarabine and Niclosamide treatment was measured by Western blotting. (B) The surface PDL1 expression level was measured by FACS with IFN-γ and Niclosamide. ( C ) The cell viability of MC38 when co-cultured with T cells and pre-treated with Fludarabine and Niclosamide were measured by CCK8 assay. ( D ) The sphere forming units induced in MC38 cells was measured with or without Niclosamide. ( E ) The cell population of CD44+/CD133+ in MC38 treated with Niclosamide was measured by FACS. ( F ) The expression of cancer stem cell marker proteins in cell nucleus was measured by Western blot. The results are expressed as the mean ± SEM of triplicate measurements in each group. *p<0.05, **p<0.01, ***p<0.001.

Article Snippet: We evaluated small molecule STAT1 inhibitor Fludarabine (F-ara-A, NSC 118218) (MedChemExpress, Cat. No.: HY-B0069) and STAT3 inhibitor Niclosamide (BAY2353, MedChemExpress, Cat. No.: HY-B0497).

Techniques: Expressing, Western Blot, Cell Culture, CCK-8 Assay, Marker

Journal: bioRxiv

Article Title: Identifying the therapeutic potential of Niclosamide in overcoming IFN-gamma dependent cancer immune evasion in the Tumor Microenvironment

doi: 10.1101/2025.10.24.684401

Figure Lengend Snippet: (A-B) The expression level of STAT1/STAT3 signaling pathway and PD-L1 proteins in MC38 cell lines with siRNA ( A ) or Fludarabine and Niclosamide treatment ( B ) was measured by Western blot. ( C ) Cell viability of MC38 cells pre-treated with different dose of IFN-γ and co-cultured with primary T cells were measured by CCK8 assay, under normoxia or hypoxia condition. ( D ) The Cell viability of MC38 cells treated with different dose of Fludarabine or Niclosamide were measured by CCK8 assay, under normoxia or hypoxia condition. The results are expressed as the mean ± SEM of measurements from 4 ∼ 6 animals per group, and each dot represents a measurement from one animal. *p<0.05, **p<0.01, ***p<0.001.

Article Snippet: We evaluated small molecule STAT1 inhibitor Fludarabine (F-ara-A, NSC 118218) (MedChemExpress, Cat. No.: HY-B0069) and STAT3 inhibitor Niclosamide (BAY2353, MedChemExpress, Cat. No.: HY-B0497).

Techniques: Expressing, Western Blot, Cell Culture, CCK-8 Assay