Journal: Neoplasia (New York, N.Y.)

Article Title: Regulation of growth, invasion and metabolism of breast ductal carcinoma through CCL2/CCR2 signaling interactions with MET receptor tyrosine kinases.

doi: 10.1016/j.neo.2022.100791

Figure Lengend Snippet: Fig. 1. CCL2 stimulation and CCR2 expression enhance MET phosphorylation through SRC dependent mechanisms in breast cancer cells. A. Flow cytometry for CCR2/MET co-expression in breast cancer cell lines. B-E. Immunoblot analysis on the effects of CCL2 treatment (100 ng/ml) of DCIS.com and HCC1937 cells (B), DCIS.com with wildtype CCR2 (WT) or CRISPR CCR2 knockout (CCR2-KO) (C), SUM225 pHAGE control or CCR2 overexpressing cells (CCR2-H) (D), and DCIS.com breast cancer cells with DMSO control with/without CCL2 and/or 10 mM PP2 (E). F. Co-immunoprecipitation analysis of DCIS.com cells treated with CCL2 for up to 30 min. G. Proximity Ligation assay for DCIS.com breast cancer cells treated with CCL2 for 5 or 15 min. Arrows point to positive signals (green fluorescence spots). DAPI overlay. Scale bar = 50 microns.

Article Snippet: For CCR2 (Biolegend cat o.357201), MET (Cell Signaling Technology cat no.9579), HK2 and KM1/2 staining, dewaxed five-micron sections were heated at low pressure n 2M urea pH 6.8 for 3 min. After quenching endogenous peroxidases with 0% methanol/3% H 2 0 2, slides were blocked in PBS/3% FBS and incubated ith primary antibodies (1:100) overnight at 4 °C.

Techniques: Expressing, Phospho-proteomics, Flow Cytometry, Western Blot, CRISPR, Knock-Out, Control, Immunoprecipitation, Proximity Ligation Assay, Fluorescence

Journal: Neoplasia (New York, N.Y.)

Article Title: Regulation of growth, invasion and metabolism of breast ductal carcinoma through CCL2/CCR2 signaling interactions with MET receptor tyrosine kinases.

doi: 10.1016/j.neo.2022.100791

Figure Lengend Snippet: Fig. 6. MET expression correlates to CCR2 expression in human breast carcinoma tissues. A. Spearman correlation test was performed for CCR2 and MET mRNA expression in IDC samples from TCGA datasets (cbioportal.org). n = 963 B. Co-immunofluorescence staining was performed in normal breast, pure DCIS, DCIS co-occurring with IDC (co-DCIS) and IDC tissues for CCR2 (red) and MET (green) expression, with DAPI counterstain, n = 9/group. Overlapping expression indicated by arrows. Scale bar = 200 microns. C. MET and CCR2 expression in DCIS and IDC tissues were quantified by Image J and normalized to DAPI. C-D. CCR2 and MET protein expression were detected in pure DCIS ( n = 33), Co-DCIS ( n = 58) (C) and IDC (D) ( n = 67) tissues by DAB immunostaining. Expression was quantified by Image J. Statistical analysis was performed using Spearman correlation test. Statistical significance was determined by p < 0.05.

Article Snippet: For CCR2 (Biolegend cat o.357201), MET (Cell Signaling Technology cat no.9579), HK2 and KM1/2 staining, dewaxed five-micron sections were heated at low pressure n 2M urea pH 6.8 for 3 min. After quenching endogenous peroxidases with 0% methanol/3% H 2 0 2, slides were blocked in PBS/3% FBS and incubated ith primary antibodies (1:100) overnight at 4 °C.

Techniques: Expressing, Immunofluorescence, Staining, Immunostaining