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NPL, DCTPP1, and PLTP significantly affected M2 macrophage polarization and CRC cells tumorigenesis (A) Overlap between M2Gs and M2 macrophage marker genes. (B) Relative expression of CTSD , TUBB , FABP5 , PLTP , NPL , and DCTPP1 in M0-like, M1-like, and M2-like macrophages. (C) <t>Immunohistochemical</t> staining of CD163, PLTP, NPL, and DCTPP1 in colorectal cancer tissues, n = 8. (D) Schematic of THP-1-derived M2-like macrophage polarization. (E and F) Proliferation and apoptosis of HCT116 cells cultured with conditioned medium from Ctrl or siNDP macrophages, n = 3. (G and H) Schematic and representative images of Transwell migration and Matrigel invasion assays. (I) Representative flow cytometry plots of CD11b and CD163 expression in macrophages under the indicated treatments, n = 3. (J and K) Ki67 immunofluorescence and quantification in HCT116 cells cultured with conditioned media from the indicated groups, n = 3. Ctrl, control siRNA; PMA-ctrl, PMA-differentiated macrophages without IL-10; siNDP, equal-molar mixture of siRNAs targeting NPL, DCTPP1, and PLTP. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001. Scale bars, 100 μm unless otherwise indicated.
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NPL, DCTPP1, and PLTP significantly affected M2 macrophage polarization and CRC cells tumorigenesis (A) Overlap between M2Gs and M2 macrophage marker genes. (B) Relative expression of CTSD , TUBB , FABP5 , PLTP , NPL , and DCTPP1 in M0-like, M1-like, and M2-like macrophages. (C) <t>Immunohistochemical</t> staining of CD163, PLTP, NPL, and DCTPP1 in colorectal cancer tissues, n = 8. (D) Schematic of THP-1-derived M2-like macrophage polarization. (E and F) Proliferation and apoptosis of HCT116 cells cultured with conditioned medium from Ctrl or siNDP macrophages, n = 3. (G and H) Schematic and representative images of Transwell migration and Matrigel invasion assays. (I) Representative flow cytometry plots of CD11b and CD163 expression in macrophages under the indicated treatments, n = 3. (J and K) Ki67 immunofluorescence and quantification in HCT116 cells cultured with conditioned media from the indicated groups, n = 3. Ctrl, control siRNA; PMA-ctrl, PMA-differentiated macrophages without IL-10; siNDP, equal-molar mixture of siRNAs targeting NPL, DCTPP1, and PLTP. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001. Scale bars, 100 μm unless otherwise indicated.
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NPL, DCTPP1, and PLTP significantly affected M2 macrophage polarization and CRC cells tumorigenesis (A) Overlap between M2Gs and M2 macrophage marker genes. (B) Relative expression of CTSD , TUBB , FABP5 , PLTP , NPL , and DCTPP1 in M0-like, M1-like, and M2-like macrophages. (C) <t>Immunohistochemical</t> staining of CD163, PLTP, NPL, and DCTPP1 in colorectal cancer tissues, n = 8. (D) Schematic of THP-1-derived M2-like macrophage polarization. (E and F) Proliferation and apoptosis of HCT116 cells cultured with conditioned medium from Ctrl or siNDP macrophages, n = 3. (G and H) Schematic and representative images of Transwell migration and Matrigel invasion assays. (I) Representative flow cytometry plots of CD11b and CD163 expression in macrophages under the indicated treatments, n = 3. (J and K) Ki67 immunofluorescence and quantification in HCT116 cells cultured with conditioned media from the indicated groups, n = 3. Ctrl, control siRNA; PMA-ctrl, PMA-differentiated macrophages without IL-10; siNDP, equal-molar mixture of siRNAs targeting NPL, DCTPP1, and PLTP. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001. Scale bars, 100 μm unless otherwise indicated.
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NPL, DCTPP1, and PLTP significantly affected M2 macrophage polarization and CRC cells tumorigenesis (A) Overlap between M2Gs and M2 macrophage marker genes. (B) Relative expression of CTSD , TUBB , FABP5 , PLTP , NPL , and DCTPP1 in M0-like, M1-like, and M2-like macrophages. (C) <t>Immunohistochemical</t> staining of CD163, PLTP, NPL, and DCTPP1 in colorectal cancer tissues, n = 8. (D) Schematic of THP-1-derived M2-like macrophage polarization. (E and F) Proliferation and apoptosis of HCT116 cells cultured with conditioned medium from Ctrl or siNDP macrophages, n = 3. (G and H) Schematic and representative images of Transwell migration and Matrigel invasion assays. (I) Representative flow cytometry plots of CD11b and CD163 expression in macrophages under the indicated treatments, n = 3. (J and K) Ki67 immunofluorescence and quantification in HCT116 cells cultured with conditioned media from the indicated groups, n = 3. Ctrl, control siRNA; PMA-ctrl, PMA-differentiated macrophages without IL-10; siNDP, equal-molar mixture of siRNAs targeting NPL, DCTPP1, and PLTP. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001. Scale bars, 100 μm unless otherwise indicated.
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NPL, DCTPP1, and PLTP significantly affected M2 macrophage polarization and CRC cells tumorigenesis (A) Overlap between M2Gs and M2 macrophage marker genes. (B) Relative expression of CTSD , TUBB , FABP5 , PLTP , NPL , and DCTPP1 in M0-like, M1-like, and M2-like macrophages. (C) <t>Immunohistochemical</t> staining of CD163, PLTP, NPL, and DCTPP1 in colorectal cancer tissues, n = 8. (D) Schematic of THP-1-derived M2-like macrophage polarization. (E and F) Proliferation and apoptosis of HCT116 cells cultured with conditioned medium from Ctrl or siNDP macrophages, n = 3. (G and H) Schematic and representative images of Transwell migration and Matrigel invasion assays. (I) Representative flow cytometry plots of CD11b and CD163 expression in macrophages under the indicated treatments, n = 3. (J and K) Ki67 immunofluorescence and quantification in HCT116 cells cultured with conditioned media from the indicated groups, n = 3. Ctrl, control siRNA; PMA-ctrl, PMA-differentiated macrophages without IL-10; siNDP, equal-molar mixture of siRNAs targeting NPL, DCTPP1, and PLTP. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001. Scale bars, 100 μm unless otherwise indicated.
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NPL, DCTPP1, and PLTP significantly affected M2 macrophage polarization and CRC cells tumorigenesis (A) Overlap between M2Gs and M2 macrophage marker genes. (B) Relative expression of CTSD , TUBB , FABP5 , PLTP , NPL , and DCTPP1 in M0-like, M1-like, and M2-like macrophages. (C) <t>Immunohistochemical</t> staining of CD163, PLTP, NPL, and DCTPP1 in colorectal cancer tissues, n = 8. (D) Schematic of THP-1-derived M2-like macrophage polarization. (E and F) Proliferation and apoptosis of HCT116 cells cultured with conditioned medium from Ctrl or siNDP macrophages, n = 3. (G and H) Schematic and representative images of Transwell migration and Matrigel invasion assays. (I) Representative flow cytometry plots of CD11b and CD163 expression in macrophages under the indicated treatments, n = 3. (J and K) Ki67 immunofluorescence and quantification in HCT116 cells cultured with conditioned media from the indicated groups, n = 3. Ctrl, control siRNA; PMA-ctrl, PMA-differentiated macrophages without IL-10; siNDP, equal-molar mixture of siRNAs targeting NPL, DCTPP1, and PLTP. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001. Scale bars, 100 μm unless otherwise indicated.
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NPL, DCTPP1, and PLTP significantly affected M2 macrophage polarization and CRC cells tumorigenesis (A) Overlap between M2Gs and M2 macrophage marker genes. (B) Relative expression of CTSD , TUBB , FABP5 , PLTP , NPL , and DCTPP1 in M0-like, M1-like, and M2-like macrophages. (C) <t>Immunohistochemical</t> staining of CD163, PLTP, NPL, and DCTPP1 in colorectal cancer tissues, n = 8. (D) Schematic of THP-1-derived M2-like macrophage polarization. (E and F) Proliferation and apoptosis of HCT116 cells cultured with conditioned medium from Ctrl or siNDP macrophages, n = 3. (G and H) Schematic and representative images of Transwell migration and Matrigel invasion assays. (I) Representative flow cytometry plots of CD11b and CD163 expression in macrophages under the indicated treatments, n = 3. (J and K) Ki67 immunofluorescence and quantification in HCT116 cells cultured with conditioned media from the indicated groups, n = 3. Ctrl, control siRNA; PMA-ctrl, PMA-differentiated macrophages without IL-10; siNDP, equal-molar mixture of siRNAs targeting NPL, DCTPP1, and PLTP. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001. Scale bars, 100 μm unless otherwise indicated.
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NPL, DCTPP1, and PLTP significantly affected M2 macrophage polarization and CRC cells tumorigenesis (A) Overlap between M2Gs and M2 macrophage marker genes. (B) Relative expression of CTSD , TUBB , FABP5 , PLTP , NPL , and DCTPP1 in M0-like, M1-like, and M2-like macrophages. (C) <t>Immunohistochemical</t> staining of CD163, PLTP, NPL, and DCTPP1 in colorectal cancer tissues, n = 8. (D) Schematic of THP-1-derived M2-like macrophage polarization. (E and F) Proliferation and apoptosis of HCT116 cells cultured with conditioned medium from Ctrl or siNDP macrophages, n = 3. (G and H) Schematic and representative images of Transwell migration and Matrigel invasion assays. (I) Representative flow cytometry plots of CD11b and CD163 expression in macrophages under the indicated treatments, n = 3. (J and K) Ki67 immunofluorescence and quantification in HCT116 cells cultured with conditioned media from the indicated groups, n = 3. Ctrl, control siRNA; PMA-ctrl, PMA-differentiated macrophages without IL-10; siNDP, equal-molar mixture of siRNAs targeting NPL, DCTPP1, and PLTP. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001. Scale bars, 100 μm unless otherwise indicated.
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NPL, DCTPP1, and PLTP significantly affected M2 macrophage polarization and CRC cells tumorigenesis (A) Overlap between M2Gs and M2 macrophage marker genes. (B) Relative expression of CTSD , TUBB , FABP5 , PLTP , NPL , and DCTPP1 in M0-like, M1-like, and M2-like macrophages. (C) <t>Immunohistochemical</t> staining of CD163, PLTP, NPL, and DCTPP1 in colorectal cancer tissues, n = 8. (D) Schematic of THP-1-derived M2-like macrophage polarization. (E and F) Proliferation and apoptosis of HCT116 cells cultured with conditioned medium from Ctrl or siNDP macrophages, n = 3. (G and H) Schematic and representative images of Transwell migration and Matrigel invasion assays. (I) Representative flow cytometry plots of CD11b and CD163 expression in macrophages under the indicated treatments, n = 3. (J and K) Ki67 immunofluorescence and quantification in HCT116 cells cultured with conditioned media from the indicated groups, n = 3. Ctrl, control siRNA; PMA-ctrl, PMA-differentiated macrophages without IL-10; siNDP, equal-molar mixture of siRNAs targeting NPL, DCTPP1, and PLTP. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001. Scale bars, 100 μm unless otherwise indicated.
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NPL, DCTPP1, and PLTP significantly affected M2 macrophage polarization and CRC cells tumorigenesis (A) Overlap between M2Gs and M2 macrophage marker genes. (B) Relative expression of CTSD , TUBB , FABP5 , PLTP , NPL , and DCTPP1 in M0-like, M1-like, and M2-like macrophages. (C) <t>Immunohistochemical</t> staining of CD163, PLTP, NPL, and DCTPP1 in colorectal cancer tissues, n = 8. (D) Schematic of THP-1-derived M2-like macrophage polarization. (E and F) Proliferation and apoptosis of HCT116 cells cultured with conditioned medium from Ctrl or siNDP macrophages, n = 3. (G and H) Schematic and representative images of Transwell migration and Matrigel invasion assays. (I) Representative flow cytometry plots of CD11b and CD163 expression in macrophages under the indicated treatments, n = 3. (J and K) Ki67 immunofluorescence and quantification in HCT116 cells cultured with conditioned media from the indicated groups, n = 3. Ctrl, control siRNA; PMA-ctrl, PMA-differentiated macrophages without IL-10; siNDP, equal-molar mixture of siRNAs targeting NPL, DCTPP1, and PLTP. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001. Scale bars, 100 μm unless otherwise indicated.
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NPL, DCTPP1, and PLTP significantly affected M2 macrophage polarization and CRC cells tumorigenesis (A) Overlap between M2Gs and M2 macrophage marker genes. (B) Relative expression of CTSD , TUBB , FABP5 , PLTP , NPL , and DCTPP1 in M0-like, M1-like, and M2-like macrophages. (C) Immunohistochemical staining of CD163, PLTP, NPL, and DCTPP1 in colorectal cancer tissues, n = 8. (D) Schematic of THP-1-derived M2-like macrophage polarization. (E and F) Proliferation and apoptosis of HCT116 cells cultured with conditioned medium from Ctrl or siNDP macrophages, n = 3. (G and H) Schematic and representative images of Transwell migration and Matrigel invasion assays. (I) Representative flow cytometry plots of CD11b and CD163 expression in macrophages under the indicated treatments, n = 3. (J and K) Ki67 immunofluorescence and quantification in HCT116 cells cultured with conditioned media from the indicated groups, n = 3. Ctrl, control siRNA; PMA-ctrl, PMA-differentiated macrophages without IL-10; siNDP, equal-molar mixture of siRNAs targeting NPL, DCTPP1, and PLTP. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001. Scale bars, 100 μm unless otherwise indicated.

Journal: iScience

Article Title: M2 macrophage associated genes shape prognosis and tumor progression in human colorectal cancer

doi: 10.1016/j.isci.2026.116230

Figure Lengend Snippet: NPL, DCTPP1, and PLTP significantly affected M2 macrophage polarization and CRC cells tumorigenesis (A) Overlap between M2Gs and M2 macrophage marker genes. (B) Relative expression of CTSD , TUBB , FABP5 , PLTP , NPL , and DCTPP1 in M0-like, M1-like, and M2-like macrophages. (C) Immunohistochemical staining of CD163, PLTP, NPL, and DCTPP1 in colorectal cancer tissues, n = 8. (D) Schematic of THP-1-derived M2-like macrophage polarization. (E and F) Proliferation and apoptosis of HCT116 cells cultured with conditioned medium from Ctrl or siNDP macrophages, n = 3. (G and H) Schematic and representative images of Transwell migration and Matrigel invasion assays. (I) Representative flow cytometry plots of CD11b and CD163 expression in macrophages under the indicated treatments, n = 3. (J and K) Ki67 immunofluorescence and quantification in HCT116 cells cultured with conditioned media from the indicated groups, n = 3. Ctrl, control siRNA; PMA-ctrl, PMA-differentiated macrophages without IL-10; siNDP, equal-molar mixture of siRNAs targeting NPL, DCTPP1, and PLTP. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001. Scale bars, 100 μm unless otherwise indicated.

Article Snippet: We paraffin-embedded colon and rectal cancer tissues and performed immunohistochemical staining following tissue sections using an Immunohistochemical Kit (Sangon Biotech, Cat# D601037) according to the manufacturer’s instructions.

Techniques: Marker, Expressing, Immunohistochemical staining, Staining, Derivative Assay, Cell Culture, Migration, Flow Cytometry, Immunofluorescence, Control