Journal: The Journal of Experimental Medicine

Article Title: Macrophages promote anti-androgen resistance in prostate cancer bone disease

doi: 10.1084/jem.20221007

Figure Lengend Snippet: Enzalutamide resistance of bone-metastatic PC can be blocked by SRC-specific inhibitor. (A) Src activity estimated using Src score (see Materials and methods) in RNA-seq data of FACS-purified MycCaP-Bo tumor cells from indicated disease stage/treatment (as described in ). (B) Src score in patient bone metastasis and metastases from other organs. (C) Correlation of Src score with macrophage abundance in patient bone metastasis datasets. (D) Correlation of Src score with ECM score in patient bone metastasis datasets. (E) Correlation of Src score with FN1 expression in patient bone metastasis datasets. (F) Correlation of Src score with ITGA5 expression in patient bone metastasis datasets. (G) Immunoblot showing the level of phosphorylated SRC (pSRC) and total SRC (t-SRC) in MycCaP-Bo cells seeded in wells precoated with 1% BSA (BSA), 1 μg/ml FN1 (FN1-1), and 10 μg/ml FN1 (FN1-10) for indicated time. (H) Immunoblotting showing the level of pSRC and t-SRC in modified MycCaP-Bo cells with doxycycline-induced expression of control shRNA or shRNA-targeting Fn1. The cells were treated with doxycycline (500 ng/ml) for 4 d. (I) Immunoblotting showing the level of pSRC and t-SRC in control MycCaP-Bo cells (Ctrl), MycCaP-Bo cells overexpressing ITGA5 clone 1 (#1) and clone 4 (#4) seeded in wells precoated with 1% BSA (BSA) or 1 μg/ml FN1 (FN1) for 6 h before sample harvest. (J) Immunoblotting showing the level of pSRC and t-SRC in in vivo MycCaP-Bo bone metastasis samples with indicated treatments. (K) Representative BLI images of resistant MycCaP-Bo bone metastasis following eCF506 treatment. (L) BLI quantification of resistant MycCaP-Bo bone metastasis following eCF506 treatment ( n = 8∼10). Data are mean ± SEM; *, P < 0.05; **, P < 0.01; ns, not significant. ANOVA was used in A and B, two-tailed unpaired Student’s t test was used for L, and Pearson correlation analysis was used in C–F. Source data are available for this figure: .

Article Snippet: After blocking in Odyssey Blocking Buffer TBS (927-50000; LI-COR Biosciences), membranes were probed with primary Abs against pSRC (D49G4, #6943; Cell Signaling Technology) or SRC (36D10, #2109; Cell Signaling Technology) overnight at 4°C.

Techniques: Activity Assay, RNA Sequencing Assay, Purification, Expressing, Western Blot, Modification, shRNA, In Vivo, Two Tailed Test

Journal: Oncotarget

Article Title: Establishment and antitumor effects of dasatinib and PKI-587 in BD-138T, a patient-derived muscle invasive bladder cancer preclinical platform with concomitant EGFR amplification and PTEN deletion

doi: 10.18632/oncotarget.10539

Figure Lengend Snippet: ( A ) Dose-response relationship curves of dasatinib and PKI-587 in 138T patient derived cell (PDC) tumors. Error bar represent the means ± SEM (triplicate). ( B ) Representative images of immunohistochemical staining of 138T PDC tumor tissue with antibodies against p-SRC (Tyr416) and p-AKT (Ser473). Scale bars, 100 μm. ( C ) 5′-ethynyl-2′-deoxyuridine (EdU) and cleaved caspase-3 analysis of 138T PDC tumors treated with dasatinib (Dose = 0, 0.5, or 1 μM) or PKI-587 (Dose = 0, 0.5, or 1 μM). Representative graphical analysis of EdU (+) and cleaved caspase-3 (+) cells following dasatinib or PKI-587 treatment; error bars represent the means ± SD. Representative images of EdU and cleaved caspase-3 staining following treatment (right).

Article Snippet: Antibodies against pSRC-Tyr416 (CST), p-p70S6K-Thr389 (CST). pAKT-Ser473 (CST), pErK(1/2)-Thr202/Tyr204 (CST), and GAPDH (CST) were used.

Techniques: Derivative Assay, Immunohistochemical staining, Staining

Journal: Oncotarget

Article Title: Establishment and antitumor effects of dasatinib and PKI-587 in BD-138T, a patient-derived muscle invasive bladder cancer preclinical platform with concomitant EGFR amplification and PTEN deletion

doi: 10.18632/oncotarget.10539

Figure Lengend Snippet: ( A ) Schematic of the establishment of PDX tumors in 138T MIBC tissues. A PDX tumor was established by subcutaneous implantation in a BALB/c-nu mouse to validate the histologic, genomic, and molecular similarity of parental and PDX tumors. ( B ) Representative images of hematoxylin & eosin (H&E) and immunohistochemical (IHC) staining of 138T PDX tumor tissues with the indicated antibodies. CK7, cytokeratin 7; pan-CK, pan-cytokeratin; CK20, cytokeratin 20. Scale bars, 100 μm (Top), 20 μm (Bottom). ( C ) Array comparative genomic hybridization (CGH) analysis of 138T parental and PDX tumors. Gene amplifications and deletions were analyzed and compared between tumors to validate genetic similarity. ( D ) Array CGH analysis of 138T PDX tumors, illustrating the mutual EGFR amplification and PTEN deletion in the PDX and parental tumors. Individual chromosome ratio plots are shown with red representing the amplified region and green representing the deleted region. High-level EGFR amplification in chromosome 7 (left). PTEN deletion in chromosome 10 (right). ( E ) EGFR amplification assessed by FISH analysis in the 138T PDX tumor. Orange signal: EGFR ; green signal: CEP7 ; blue signal: DAPI counterstaining. ( F ) Representative images of IHC staining of 138T PDX tumor tissue with EGFR, p-EGFR (Tyr1068), PTEN, p-SRC (Tyr416), and p-AKT (Ser473) antibodies. Scale bars, 100 μm.

Article Snippet: Antibodies against pSRC-Tyr416 (CST), p-p70S6K-Thr389 (CST). pAKT-Ser473 (CST), pErK(1/2)-Thr202/Tyr204 (CST), and GAPDH (CST) were used.

Techniques: Immunohistochemical staining, Immunohistochemistry, Hybridization, Amplification

Journal: Oncotarget

Article Title: Establishment and antitumor effects of dasatinib and PKI-587 in BD-138T, a patient-derived muscle invasive bladder cancer preclinical platform with concomitant EGFR amplification and PTEN deletion

doi: 10.18632/oncotarget.10539

Figure Lengend Snippet: ( A ) Measurement of subcutaneous xenograft tumor size after treatment with dasatinib or PKI-587. ** P < 0.01. *** P < 0.001. PDX tumors were treated with dasatinib (15 mg/kg, n = 8) and PKI-587 (25 mg/kg, n = 8) for 17 days, harvested, and processed for immunoblots. ( B ) Measurement of mouse body weight after treatment with dasatinib or PKI-587 showing no toxicity from the drug therapy in the PDX mouse model. Body weights were measured after treatment with dasatinib or PKI-587 for 17 days. ( C ) Immunoblots of p-SRC (Tyr416), p-p70S6K (Thr389) and p-AKT (Ser473) in xenograft tumors from (A). ( D ) Cell proliferation and apoptosis were measured immunohistochemically (IHC) by performing Ki-67 and TUNEL staining using PDX tissues treated with dasatinib or PKI-587 (left). Scale bar, 100 μm. Representative bar graphs of the IHC images (right), ** P < 0.01, *** P < 0.001 ( n = 5 fields; means ± SD, analyzed by two-tailed paired t -test).

Article Snippet: Antibodies against pSRC-Tyr416 (CST), p-p70S6K-Thr389 (CST). pAKT-Ser473 (CST), pErK(1/2)-Thr202/Tyr204 (CST), and GAPDH (CST) were used.

Techniques: Western Blot, TUNEL Assay, Staining, Two Tailed Test