Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Breast cancer patient-derived microtumors resemble tumor heterogeneity and enable protein-based stratification and functional validation of individualized drug treatment

doi: 10.1186/s13046-023-02782-2

Figure Lengend Snippet: Antibodies for IHC staining

Article Snippet: rabbit anti-human FAPalpha , BioRad , AHP1322 , Rb Linker , 1:50.

Techniques:

Journal: JCI Insight

Article Title: CSF1/CSF1R signaling mediates malignant pleural effusion formation

doi: 10.1172/jci.insight.155300

Figure Lengend Snippet: ( A and B ) CAFs in tumors of vehicle- and CSF1Ri-treated animals were visualized upon FAPA staining. ( A ) Representative pictures. Scale bar: 50 μm. ( B ) Results of image analysis. Data presented as mean ± SEM. LLC: n = 4 for both groups. MC38: n = 5 for both groups. * P < 0.05 compared with vehicle by 2-tailed Student’s t test. HPF, high-power field. ( C ) Activated CAFs isolated from LLC or MC38 tumors were treated with vehicle or 6700 nM CSF1Ri overnight and subsequently loaded with 7 × 10 4 tumor cells/well; 8 hours later 2 × 10 5 M1 macrophages/well were added to the tumor cell–CAF cocultures; 24 hours later, tumor cells were counted. ( D and E ) CD45 + CD11b + F4/80 + macrophages were analyzed for IL-10 ( D ) and IL-12 ( E ) expression. Data presented as mean ± SEM, 2 independent experiments. LLC: vehicle-treated CAFs n = 5, CSF1Ri-treated CAFs n = 5. MC38: vehicle-treated CAFs n = 7, CSF1Ri-treated CAFs n = 7. * P < 0.05 compared with vehicle-treated CAFs by 2-tailed Student’s t test. ( F and G ) Isolated CAFs from LLC and MC38 tumors (treated with vehicle or CSF1Ri overnight) were cocultured with macrophages for 48 hours. Macrophages were isolated and mRNA levels of Vegfa ( F ) and Il6 ( G ) were quantified by real-time PCR. Data presented as mean ± SEM. n = 7 for both groups, * P < 0.05 compared with indicated group by 2-tailed Student’s t test. ( H ) Isolated CAFs from LLC or MC38 tumors were treated with vehicle, CSF1Ri, CSF1, or IL-34 or pretreated with CSF1Ri for 2 hours and then CSF1 was added; 24 hours later, MIP2 levels were quantified in supernatants. Results were normalized to total protein. Data presented as mean ± SEM. Vehicle n = 6, CSF1Ri n = 6, CSF1 n = 5, CSF1Ri+CSF1 n = 4, IL-34 n = 5 (3 independent experiments). * P < 0.05 compared with indicated group by 1-way ANOVA (with Tukey’s post hoc test).

Article Snippet: CAFs were isolated from LLC or MC38 tumors by FAPA antibody–loaded (CSB-PA008191, Cusabio Technology) magnetic beads (MojoSort Streptavidin Nanobeads, BioLegend) as previously reported ( ).

Techniques: Staining, Isolation, Expressing, Real-time Polymerase Chain Reaction

Journal: Cancers

Article Title: NAB 2-Expressing Cancer-Associated Fibroblast Promotes HNSCC Progression

doi: 10.3390/cancers11030388

Figure Lengend Snippet: CAF marker and matrix metalloproteinase (MMP) expression in primary fibroblasts from HNSCC patient tissue. ( A ) CAFs and paired non-tumor fibroblasts (NTFs) were isolated from three HNSCC patient tissue (P1, P2, P3) specimens by short-term primary culture. α-SMA and FAP mRNA and protein levels were evaluated by real-time quantitative polymerase chain reaction (qPCR) and Western blotting, respectively. ( B ) MMP mRNA and protein expression was analyzed under the same conditions. Results represent the mean ± standard deviation of two or three experiments.

Article Snippet: Rabbit anti-fibroblast activation protein (FAP) antibody was from GeneTex (GeneTex, Inc., Irvine, CA, USA).

Techniques: Marker, Expressing, Isolation, Real-time Polymerase Chain Reaction, Western Blot, Standard Deviation