Journal: Cancers

Article Title: Drug-Induced Resistance and Phenotypic Switch in Triple-Negative Breast Cancer Can Be Controlled via Resolution and Targeting of Individualized Signaling Signatures

doi: 10.3390/cancers13195009

Figure Lengend Snippet: TNBC tissues are represented by different patient-specific signaling signatures, majority of which do not include EGFR. ( A ) Fold changes in expression levels of EGFR and pEGFR in TNBC and non-TNBC tumors are shown. Y1068 and Y1173 are EGFR phosphorylation sites; ( B ) Examples for patient-specific sets of active unbalanced processes are shown. Each sample harbors a set of 1–3 active unbalanced processes (PaSSS), represented schematically by a barcode. In each barcode active unbalanced processes are represented by black or gray squares, inactive white. Negative/positive amplitude denotes how the patients are correlated with respect to a particular process. Suggested PaSSS-based therapies appear below each barcode; ( C ) Heterogeneity index of 3 subgroups, represented by a ratio between the number of distinct PaSSSs and the number of samples in each subset, is shown for the TNBC subset of tissues, the entire set (3467 samples from 11 cancer types) and the subset of non-TNBC samples. (Abbreviations: TNBC—Triple Negative Breast Cancer, PaSSS—Patient-specific signaling signature, EGFR—Epidermal Growth Factor Receptor, VEGFR2—Vascular Endothelial Growth Factor Receptor 2, Her2—Human Epidermal growth factor Receptor 2, Src—Proto-oncogene tyrosine-protein kinase Src).

Article Snippet: The following conjugated antibodies were used: anti-p-EGFR (Y1068) (R&D Systems, Minneapolis, MN, USA, cat. no. IC3570G), anti-p-ERK2 (Thr202/Tyr204) (BioLegend, San Diego, CA, USA, cat. No. 675503), anti-p-S6 (Ser235/236) (BioLegend, cat. no. 608605), and anti-GAPDH (Santa Cruz Biotechnology, Dallas, Texas, USA, cat. no. sc-47724AF594).

Techniques: Expressing, Phospho-proteomics

Journal: JCI Insight

Article Title: ADAM17 substrate release in proximal tubule drives kidney fibrosis

doi: 10.1172/jci.insight.87023

Figure Lengend Snippet: The expression levels of ADAM17 and its pathway components were examined at different time points after ischemic injury in whole kidneys. (A) qPCR analysis of ADAM17 mRNA expression levels in ex/ex or WT/WT mice, expressed as fold over respective sham-injured mice (n = 3–5). (B) Western blot analysis of ADAM17 protein levels in ex/ex or WT/WT mice subjected to sham or IRI (day 5 after ischemia) (left: sample blots; right: quantification; GAPDH was used as loading control; n = 5). (C–H) qPCR analysis of EGFR ligands, EGFR, TNFα, and TNFR1/2 mRNA expression levels in ex/ex or WT/WT mice, expressed as fold over respective sham-injured mice (time points as indicated; n = 3–6). (I) Soluble ectodomains of the ADAM17 substrates TNFα, TGFα, and AREG were measured in whole kidney lysates of WT/WT or ex/ex mice at day 2 after ischemia by ELISA (ADAM10 substrate cMET is used as control; n = 4). (J) Time-course of EGFR phosphorylation (Y1068) as measured by Western blot in whole kidney lysates of WT/WT or ex/ex mice (n = 5). (K) Representative images of EGFR phosphorylation (Y1173) examined in kidney cortex by immunostaining in WT/WT or ex/ex mice at day 2 after injury (left: representative images; right: quantification; n = 3; scale bar: 50 μm). *P < 0.05; **P < 0.01; ***P < 0.001 as determined by an unpaired 2-tailed Student’s t test. ADAM17; a disintegrin and metalloprotease 17; EGFR, epidermal growth factor receptor; IRI, ischemia reperfusion injury; NS, nonspecific band; TNFα, tumor necrosis factor α; TNFR, tumor necrosis factor receptor; AREG, amphiregulin; ADAM10, a disintegrin and metalloprotease 10; cMet, met proto-oncogene; WB, Western blot.

Article Snippet: The primary Abs included: ADAM17 (a gift from Carl Blobel, Hospital for Special Surgery, New York, New York, USA), αSMA (CY3-conjugated, Sigma-Aldrich, C6198), fibronectin (Abcam, ab2413), CD31 (eBioscience, 14–0311) pEGFR Y1068 (Cell Signaling Technology, 3777), pEGFR Y1173 (R&D Systems, AF1095), and AREG (R&D Systems, BAF262).

Techniques: Expressing, Western Blot, Control, Enzyme-linked Immunosorbent Assay, Phospho-proteomics, Immunostaining