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(A) Schematic of TAM extraction from murine TNBC tumors. (B) Flow cytometry analysis gating strategy comparing CXCL9 protein abundance captured in the media. (C) Flow cytometry analysis gating for CD11b+F4/80+ cells after culture. (D) Bar plots summarize flow cytometry across replicates for CXCL9 abundance in media (T-test, p < 0.001, n=3, SEM bars) (E) Bar plots summarize flow cytometry across replicates for CXCL9 and iNOS within TAMs. (One way ANOVA, p < 0.001 for all comparisons, n=6, SEM bars) (F) Schematic of TAM expression of markers after reprogramming. (G) Western blot analysis comparing STAT, MAPK, and TAM marker expression changes across in vitro treatments in 2153L TAMs. (H) Western blot analysis comparing STAT, MAPK, and TAM marker expression changes across in vitro treatments in T12 TAMs. (I) Dot plot for Day 7 scRNAseq dataset showing <t>STAT1</t> and SOCS1 expression enriched in the CXCL9+ Mo.Mac population. Created in BioRender. Zhao, N. (2026) https://BioRender.com/e6uqj0s .
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(A) Schematic of TAM extraction from murine TNBC tumors. (B) Flow cytometry analysis gating strategy comparing CXCL9 protein abundance captured in the media. (C) Flow cytometry analysis gating for CD11b+F4/80+ cells after culture. (D) Bar plots summarize flow cytometry across replicates for CXCL9 abundance in media (T-test, p < 0.001, n=3, SEM bars) (E) Bar plots summarize flow cytometry across replicates for CXCL9 and iNOS within TAMs. (One way ANOVA, p < 0.001 for all comparisons, n=6, SEM bars) (F) Schematic of TAM expression of markers after reprogramming. (G) Western blot analysis comparing STAT, MAPK, and TAM marker expression changes across in vitro treatments in 2153L TAMs. (H) Western blot analysis comparing STAT, MAPK, and TAM marker expression changes across in vitro treatments in T12 TAMs. (I) Dot plot for Day 7 scRNAseq dataset showing <t>STAT1</t> and SOCS1 expression enriched in the CXCL9+ Mo.Mac population. Created in BioRender. Zhao, N. (2026) https://BioRender.com/e6uqj0s .
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(A) Schematic of TAM extraction from murine TNBC tumors. (B) Flow cytometry analysis gating strategy comparing CXCL9 protein abundance captured in the media. (C) Flow cytometry analysis gating for CD11b+F4/80+ cells after culture. (D) Bar plots summarize flow cytometry across replicates for CXCL9 abundance in media (T-test, p < 0.001, n=3, SEM bars) (E) Bar plots summarize flow cytometry across replicates for CXCL9 and iNOS within TAMs. (One way ANOVA, p < 0.001 for all comparisons, n=6, SEM bars) (F) Schematic of TAM expression of markers after reprogramming. (G) Western blot analysis comparing STAT, MAPK, and TAM marker expression changes across in vitro treatments in 2153L TAMs. (H) Western blot analysis comparing STAT, MAPK, and TAM marker expression changes across in vitro treatments in T12 TAMs. (I) Dot plot for Day 7 scRNAseq dataset showing <t>STAT1</t> and SOCS1 expression enriched in the CXCL9+ Mo.Mac population. Created in BioRender. Zhao, N. (2026) https://BioRender.com/e6uqj0s .
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(A) Schematic of TAM extraction from murine TNBC tumors. (B) Flow cytometry analysis gating strategy comparing CXCL9 protein abundance captured in the media. (C) Flow cytometry analysis gating for CD11b+F4/80+ cells after culture. (D) Bar plots summarize flow cytometry across replicates for CXCL9 abundance in media (T-test, p < 0.001, n=3, SEM bars) (E) Bar plots summarize flow cytometry across replicates for CXCL9 and iNOS within TAMs. (One way ANOVA, p < 0.001 for all comparisons, n=6, SEM bars) (F) Schematic of TAM expression of markers after reprogramming. (G) Western blot analysis comparing STAT, MAPK, and TAM marker expression changes across in vitro treatments in 2153L TAMs. (H) Western blot analysis comparing STAT, MAPK, and TAM marker expression changes across in vitro treatments in T12 TAMs. (I) Dot plot for Day 7 scRNAseq dataset showing <t>STAT1</t> and SOCS1 expression enriched in the CXCL9+ Mo.Mac population. Created in BioRender. Zhao, N. (2026) https://BioRender.com/e6uqj0s .
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(A) Schematic of TAM extraction from murine TNBC tumors. (B) Flow cytometry analysis gating strategy comparing CXCL9 protein abundance captured in the media. (C) Flow cytometry analysis gating for CD11b+F4/80+ cells after culture. (D) Bar plots summarize flow cytometry across replicates for CXCL9 abundance in media (T-test, p < 0.001, n=3, SEM bars) (E) Bar plots summarize flow cytometry across replicates for CXCL9 and iNOS within TAMs. (One way ANOVA, p < 0.001 for all comparisons, n=6, SEM bars) (F) Schematic of TAM expression of markers after reprogramming. (G) Western blot analysis comparing STAT, MAPK, and TAM marker expression changes across in vitro treatments in 2153L TAMs. (H) Western blot analysis comparing STAT, MAPK, and TAM marker expression changes across in vitro treatments in T12 TAMs. (I) Dot plot for Day 7 scRNAseq dataset showing <t>STAT1</t> and SOCS1 expression enriched in the CXCL9+ Mo.Mac population. Created in BioRender. Zhao, N. (2026) https://BioRender.com/e6uqj0s .
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(A) Schematic of TAM extraction from murine TNBC tumors. (B) Flow cytometry analysis gating strategy comparing CXCL9 protein abundance captured in the media. (C) Flow cytometry analysis gating for CD11b+F4/80+ cells after culture. (D) Bar plots summarize flow cytometry across replicates for CXCL9 abundance in media (T-test, p < 0.001, n=3, SEM bars) (E) Bar plots summarize flow cytometry across replicates for CXCL9 and iNOS within TAMs. (One way ANOVA, p < 0.001 for all comparisons, n=6, SEM bars) (F) Schematic of TAM expression of markers after reprogramming. (G) Western blot analysis comparing STAT, MAPK, and TAM marker expression changes across in vitro treatments in 2153L TAMs. (H) Western blot analysis comparing STAT, MAPK, and TAM marker expression changes across in vitro treatments in T12 TAMs. (I) Dot plot for Day 7 scRNAseq dataset showing <t>STAT1</t> and SOCS1 expression enriched in the CXCL9+ Mo.Mac population. Created in BioRender. Zhao, N. (2026) https://BioRender.com/e6uqj0s .
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(A) Schematic of TAM extraction from murine TNBC tumors. (B) Flow cytometry analysis gating strategy comparing CXCL9 protein abundance captured in the media. (C) Flow cytometry analysis gating for CD11b+F4/80+ cells after culture. (D) Bar plots summarize flow cytometry across replicates for CXCL9 abundance in media (T-test, p < 0.001, n=3, SEM bars) (E) Bar plots summarize flow cytometry across replicates for CXCL9 and iNOS within TAMs. (One way ANOVA, p < 0.001 for all comparisons, n=6, SEM bars) (F) Schematic of TAM expression of markers after reprogramming. (G) Western blot analysis comparing STAT, MAPK, and TAM marker expression changes across in vitro treatments in 2153L TAMs. (H) Western blot analysis comparing STAT, MAPK, and TAM marker expression changes across in vitro treatments in T12 TAMs. (I) Dot plot for Day 7 scRNAseq dataset showing <t>STAT1</t> and SOCS1 expression enriched in the CXCL9+ Mo.Mac population. Created in BioRender. Zhao, N. (2026) https://BioRender.com/e6uqj0s .
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(A) Schematic of TAM extraction from murine TNBC tumors. (B) Flow cytometry analysis gating strategy comparing CXCL9 protein abundance captured in the media. (C) Flow cytometry analysis gating for CD11b+F4/80+ cells after culture. (D) Bar plots summarize flow cytometry across replicates for CXCL9 abundance in media (T-test, p < 0.001, n=3, SEM bars) (E) Bar plots summarize flow cytometry across replicates for CXCL9 and iNOS within TAMs. (One way ANOVA, p < 0.001 for all comparisons, n=6, SEM bars) (F) Schematic of TAM expression of markers after reprogramming. (G) Western blot analysis comparing STAT, MAPK, and TAM marker expression changes across in vitro treatments in 2153L TAMs. (H) Western blot analysis comparing STAT, MAPK, and TAM marker expression changes across in vitro treatments in T12 TAMs. (I) Dot plot for Day 7 scRNAseq dataset showing STAT1 and SOCS1 expression enriched in the CXCL9+ Mo.Mac population. Created in BioRender. Zhao, N. (2026) https://BioRender.com/e6uqj0s .

Journal: Cancer immunology research

Article Title: MERTK inhibition cooperates with immunomodulatory cyclophosphamide to induce CXCL9 + monocyte-macrophage programming and durable antitumor immunity in triple negative breast cancer

doi: 10.1158/2326-6066.CIR-26-0278

Figure Lengend Snippet: (A) Schematic of TAM extraction from murine TNBC tumors. (B) Flow cytometry analysis gating strategy comparing CXCL9 protein abundance captured in the media. (C) Flow cytometry analysis gating for CD11b+F4/80+ cells after culture. (D) Bar plots summarize flow cytometry across replicates for CXCL9 abundance in media (T-test, p < 0.001, n=3, SEM bars) (E) Bar plots summarize flow cytometry across replicates for CXCL9 and iNOS within TAMs. (One way ANOVA, p < 0.001 for all comparisons, n=6, SEM bars) (F) Schematic of TAM expression of markers after reprogramming. (G) Western blot analysis comparing STAT, MAPK, and TAM marker expression changes across in vitro treatments in 2153L TAMs. (H) Western blot analysis comparing STAT, MAPK, and TAM marker expression changes across in vitro treatments in T12 TAMs. (I) Dot plot for Day 7 scRNAseq dataset showing STAT1 and SOCS1 expression enriched in the CXCL9+ Mo.Mac population. Created in BioRender. Zhao, N. (2026) https://BioRender.com/e6uqj0s .

Article Snippet: Blots were blocked with 5% non-fat dry milk (BioRad, 1706404) in PBS, then probed with the following antibodies in 5% BSA (Sigma-Aldrich, A7906) solution: MERTK (Cell Signaling Technology (CST), 38102), AXL (CST, 8661, RRID:AB_11217435), p44/42 (CST, 9102, RRID:AB_330744), p-p44/42 (CST, 9101, RRID:AB_331646), iNOS (CST, 13120, RRID:AB_2799268), PD-L1 (R&D Systems, AF1019, RRID:AB_354540), p-STAT1 (CST, 8062, RRID:AB_10949108), Arg1(CST, 93668, RRID:AB_2800207), and SOCS1 (CST, 55313).

Techniques: Inhibition, Extraction, Flow Cytometry, Analysis, Quantitative Proteomics, Expressing, Western Blot, Marker, In Vitro