Journal: Cell Communication and Signaling : CCS

Article Title: Aurantio-obtusin modulates Wilms Tumour 1 within the breast tumour microenvironment reducing immunosuppression and tumour growth

doi: 10.1186/s12964-025-02292-y

Figure Lengend Snippet: WT1 regulates the STAT1/3 pathway in CAFs. A Protein levels of WT1, STAT1, STAT3, PD-L1 and the TGFβ receptor were determined in WT1 knockdown ( A ) and WT1 overexpressed ( B ) CAFs by Western blotting. Actin was used as a loading control. IDO release was determined by ELISA. RNA levels of PD-L1 was determined by qRT-PCR. TBP was used as a loading control. Representative experiment is shown of n = 3–5 biological replicates. Data represents mean ± SEM; * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001

Article Snippet: The blots were probed with primary antibodies against WT1 (Invitrogen, MA5-38,406; 1:500), STAT1 (Invitrogen, AHO0832; 1:500), STAT3 (Invitrogen, PA5-85,199; 1:500), PD-L1 (R&D systems, AF156; 1:100), TGF-β RI (R&D systems, AF3025; 1:500), p53 (R&D systems, AF1355; 1:1,000), β-actin (Sigma Aldrich, A5441; 1:20,000), and GAPDH (Cell Signaling Technology, 2118S; 1:1,000) overnight at 4 °C and detected by chemiluminescence using Goat anti-Rabbit IgG secondary antibody (Invitrogen, 31,460; 1:5,000), Rabbit anti-Goat IgG secondary antibody (Invitrogen, 31,402; 1:5,000), Goat anti-mouse IgG secondary antibody (Invitrogen, 31,430; 1:5,000).

Techniques: Knockdown, Western Blot, Control, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR

Journal: Cell Communication and Signaling : CCS

Article Title: Aurantio-obtusin modulates Wilms Tumour 1 within the breast tumour microenvironment reducing immunosuppression and tumour growth

doi: 10.1186/s12964-025-02292-y

Figure Lengend Snippet: Chemotherapeutic drugs DOX and PTX increase WT1 levels in CAFs and enhance their ability to inhibit T cell proliferation. A Schematic overview of drug treatment, co-culture of CAFs and PBMCs followed by T cell proliferation assay. B Protein expression of WT1 and p53 in CAFs following DOX and PTX treatment was determined by Western blotting. GAPDH was used as a loading control. C Protein levels of STAT1, STAT3, PD-L1 levels. IDO release from DOX and PTX treated CAFs was determined by ELISA. D DOX treated CAFs were co-cultured with PBMCs and CFSE-labelled CD4 + and CD8 + T cell proliferation was determined by flow cytometry ( E ) PTX treated CAFs were co-cultured with PBMCs and CFSE-labelled CD4 + and CD8. + T cell proliferation was determined by flow cytometry. Representative experiment is shown of n = 3 biological replicates. Data are mean ± SEM; * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001

Article Snippet: The blots were probed with primary antibodies against WT1 (Invitrogen, MA5-38,406; 1:500), STAT1 (Invitrogen, AHO0832; 1:500), STAT3 (Invitrogen, PA5-85,199; 1:500), PD-L1 (R&D systems, AF156; 1:100), TGF-β RI (R&D systems, AF3025; 1:500), p53 (R&D systems, AF1355; 1:1,000), β-actin (Sigma Aldrich, A5441; 1:20,000), and GAPDH (Cell Signaling Technology, 2118S; 1:1,000) overnight at 4 °C and detected by chemiluminescence using Goat anti-Rabbit IgG secondary antibody (Invitrogen, 31,460; 1:5,000), Rabbit anti-Goat IgG secondary antibody (Invitrogen, 31,402; 1:5,000), Goat anti-mouse IgG secondary antibody (Invitrogen, 31,430; 1:5,000).

Techniques: Co-Culture Assay, Proliferation Assay, Expressing, Western Blot, Control, Enzyme-linked Immunosorbent Assay, Cell Culture, Flow Cytometry

Journal: Cell Communication and Signaling : CCS

Article Title: Aurantio-obtusin modulates Wilms Tumour 1 within the breast tumour microenvironment reducing immunosuppression and tumour growth

doi: 10.1186/s12964-025-02292-y

Figure Lengend Snippet: AO reduces WT1 levels in CAFs and reduces the ability of CAFs to suppress T cell proliferation. A Comparison of chemical structures of DOX and AO. B Protein levels of WT1, STAT1, STAT3, and PD-L1 after AO treatment of CAFs were determined by Western blotting. Actin was used as a loading control. C Dose–response curves of DOX, PTX and AO treatment of CAFs. P1 and P3 (Luminal A), P2 (TNBC). Curves were generated by non-linear regression analysis by GraphPad. D AO treated CAFs were co-cultured with PBMCs and CFSE-labelled CD4 + and CD8. + T cell proliferation was determined by flow cytometry. Representative experiment is shown of n = 3 biological replicates. Data represent mean ± SEM; * P < 0.05, and ** P < 0.01

Article Snippet: The blots were probed with primary antibodies against WT1 (Invitrogen, MA5-38,406; 1:500), STAT1 (Invitrogen, AHO0832; 1:500), STAT3 (Invitrogen, PA5-85,199; 1:500), PD-L1 (R&D systems, AF156; 1:100), TGF-β RI (R&D systems, AF3025; 1:500), p53 (R&D systems, AF1355; 1:1,000), β-actin (Sigma Aldrich, A5441; 1:20,000), and GAPDH (Cell Signaling Technology, 2118S; 1:1,000) overnight at 4 °C and detected by chemiluminescence using Goat anti-Rabbit IgG secondary antibody (Invitrogen, 31,460; 1:5,000), Rabbit anti-Goat IgG secondary antibody (Invitrogen, 31,402; 1:5,000), Goat anti-mouse IgG secondary antibody (Invitrogen, 31,430; 1:5,000).

Techniques: Comparison, Western Blot, Control, Generated, Cell Culture, Flow Cytometry

Journal: Cell Communication and Signaling : CCS

Article Title: Aurantio-obtusin modulates Wilms Tumour 1 within the breast tumour microenvironment reducing immunosuppression and tumour growth

doi: 10.1186/s12964-025-02292-y

Figure Lengend Snippet: Proposed model highlighting how WT1 in CAFs modulates immune evasion and tumour growth in the breast TME. WT1 is upregulated in breast patient-derived CAFs. WT1 is a transcription factor, regulating expression of STAT1/STAT3 in CAFs. STAT1/STAT3 regulate PD-L1 expression and IDO release in CAFs. Both PD-L1 and IDO inhibit T cell activity and cytotoxic GZMB levels within the TME, contributing to increased tumour growth. Targeting WT1 in CAFs reduces the release of immunosuppressive factors into the TME. T cells are more active and are able to limit tumour growth

Article Snippet: The blots were probed with primary antibodies against WT1 (Invitrogen, MA5-38,406; 1:500), STAT1 (Invitrogen, AHO0832; 1:500), STAT3 (Invitrogen, PA5-85,199; 1:500), PD-L1 (R&D systems, AF156; 1:100), TGF-β RI (R&D systems, AF3025; 1:500), p53 (R&D systems, AF1355; 1:1,000), β-actin (Sigma Aldrich, A5441; 1:20,000), and GAPDH (Cell Signaling Technology, 2118S; 1:1,000) overnight at 4 °C and detected by chemiluminescence using Goat anti-Rabbit IgG secondary antibody (Invitrogen, 31,460; 1:5,000), Rabbit anti-Goat IgG secondary antibody (Invitrogen, 31,402; 1:5,000), Goat anti-mouse IgG secondary antibody (Invitrogen, 31,430; 1:5,000).

Techniques: Derivative Assay, Expressing, Activity Assay

Journal: Frontiers in Medicine

Article Title: Serum sodium levels associate with recovery of kidney function in immune checkpoint inhibitor nephrotoxicity

doi: 10.3389/fmed.2023.1020691

Figure Lengend Snippet: Clinical, laboratory, and histopathological parameters of the total cohort.

Article Snippet: After deparaffinization in xylene and rehydration in ethanol containing distilled water, formalin-fixed, paraffin-embedded kidney sections were stained using primary antibodies against PD-L1 (1:100, ab205921, Abcam, Cambridge, UK), PD-1 (1:500, ab52587, Abcam, Cambridge, UK), C1q (1:30,000, A0136, Agilent Dako, Santa Clara, USA), and C3c (1:10,000, A0062, Agilent Dako, Santa Clara, USA), labeling was performed using NovolinkTM Polymer Detection System (Leica Biosystems, Wetzlar, Germany) according to the manufacturer’s protocol.

Techniques:

Journal: Frontiers in Medicine

Article Title: Serum sodium levels associate with recovery of kidney function in immune checkpoint inhibitor nephrotoxicity

doi: 10.3389/fmed.2023.1020691

Figure Lengend Snippet: Histopathological lesions or cumulative steroid dose do not affect renal recovery after AIN related to ICI nephrotoxicity. (A) Difference between worst eGFR at time of diagnosis and last documented eGFR during follow-up in the patient cohort are shown. (B) Intensity of PD-L1/PD-1 positivity within different renal compartments, cumulative steroid dose and histopathological findings in association with eGFR recovery in AIN related to ICI nephrotoxicity are shown by heatmap reflecting mean values of Spearman’s ρ. AIN, acute interstitial nephritis; eGFR, estimated glomerular filtration rate (CKD-EPI); ICI, immune checkpoint inhibitor; IF/TA, interstitial fibrosis/tubular atrophy; PD-1, programmed cell death protein 1; PD-L1, programmed cell death protein 1-ligand 1.

Article Snippet: After deparaffinization in xylene and rehydration in ethanol containing distilled water, formalin-fixed, paraffin-embedded kidney sections were stained using primary antibodies against PD-L1 (1:100, ab205921, Abcam, Cambridge, UK), PD-1 (1:500, ab52587, Abcam, Cambridge, UK), C1q (1:30,000, A0136, Agilent Dako, Santa Clara, USA), and C3c (1:10,000, A0062, Agilent Dako, Santa Clara, USA), labeling was performed using NovolinkTM Polymer Detection System (Leica Biosystems, Wetzlar, Germany) according to the manufacturer’s protocol.

Techniques: Biomarker Discovery, Filtration