Journal: Cellular and Molecular Immunology

Article Title: Reprogramming of cancer-associated fibroblasts by apoptotic cancer cells inhibits lung metastasis via Notch1-WISP-1 signaling

doi: 10.1038/s41423-022-00930-w

Figure Lengend Snippet: WISP-1 is required for the antimigratory and anti-invasive effects. a Cytokine array analysis of CAF CM, ApoSQ-CAF CM, or ApoSQ CM for 24 h. b Relative image densities of cytokine spots for ApoSQ-CAF CM and ApoSQ CM compared with CAF CM. c Immunoblot analysis of WISP-1 in CAFs transfected with WISP-1 siRNA ( upper ). Densitometric analysis of the relative WISP-1 abundance ( lower ). d ELISA of WISP-1 in CM from CAFs transfected with WISP-1 siRNA for 24 h before treatment with or without ApoSQ for 20 h. Quantification of migrated ( e ) and invaded ( f ) 344SQ cells. e , f CAFs were transfected with control or WISP-1 siRNA before exposure to ApoSQ for 20 h. CM was added to 344SQ cells with TGF-β1 (10 ng/ml) for 48 h. Quantification of migrated ( g ) and invaded ( h ) CAFs. g , h CAFs were transfected with WISP-1 siRNA before exposure to ApoSQ for 20 h, which the medium was then replaced with fresh medium containing TGF-β1 (10 ng/ml) for 24 h. i Phase contrast microscopy ( left ) and quantification of migrated and invaded 344SQ cells ( right) . CAF CM and ApoSQ-CAF CM were treated with a neutralizing anti-WISP-1 Ab (10 μg/ml) or IgG isotype control before addition to 344SQ cells with or without TGF-β1 (10 ng/ml) for 48 h. j Immunoblot analysis of WISP-1 in CAFs transfected with control vector (mock) or the WISP-1 plasmid (WISP-1 O/E) for 24 h ( upper ). Densitometric analysis of the relative WISP-1 protein abundance ( lower ). k ELISA of WISP-1 in CM from CAFs transfected with empty vector or the WISP-1 plasmid for 24 h before treatment with or without ApoSQ for 20 h. l Phase contrast microscopy ( left ) and quantification of migrated and invaded 344SQ cells ( right ). CM from CAFs transfected with empty vector or the WISP-1 plasmid before treatment with or without ApoSQ for 20 h was added to 344SQ cells with TGF-β1 (10 ng/ml) for 48 h. i , l Scale bars: 100 μm. NS, NS: not significant, * P < 0.05, ** P < 0.01, *** P < 0.001, two-tailed Student’s t test. The data are from one experiment representative of three independent experiments with similar results ( c and j upper , i and l left ) or from three independent experiments (mean ± standard error; c and j lower , d – h , i and l right , k )

Article Snippet: Gene overexpression experiments were performed with CAFs seeded at 0.5 × 10 6 cells/ml in 60 mm dishes for 24 h. For WISP-1 overexpression, cells were transfected with 2.5 μg of the mouse WISP-1 gene ORF cRNA clone expression plasmid (MG5A0255-CH, Sino Biological, North Wales, PA, USA) using Lipofectamine 2000 (Thermo Fisher Scientific, Waltham, MA, USA) for 24 h according to the manufacturer’s instructions.

Techniques: Western Blot, Transfection, Enzyme-linked Immunosorbent Assay, Microscopy, Plasmid Preparation, Two Tailed Test

Journal: Cellular and Molecular Immunology

Article Title: Reprogramming of cancer-associated fibroblasts by apoptotic cancer cells inhibits lung metastasis via Notch1-WISP-1 signaling

doi: 10.1038/s41423-022-00930-w

Figure Lengend Snippet: WISP-1 acts through integrin αvβ3 in 344SQ cells and αvβ5 in CAFs to inhibit TGF-β1-induced migration and invasion. a Phase contrast microscopy ( left ) and quantification of migrated and invaded 344SQ cells ( right) . 344SQ cells were pretreated with an anti-integrin blocking antibody (3 μg/ml; anti-integrin αν, α5, β1, β3, or β5) or corresponding IgG isotype control for 30 min before treatment with rWISP-1 (10 ng/ml) and TGF-β1 (10 ng/ml) for 24 h. b Phase contrast microscopy ( left ) and quantification of migrated and invaded CAFs ( right) . CAFs were pretreated with an anti-integrin blocking antibody (10 μg/ml) for 1 h before treatment with rWISP-1 (3 ng/ml) and TGF-β1 (10 ng/ml) for 24 h. Scale bars: 100 μm. Immunoblot analysis of the indicated proteins in 344SQ cell ( c , e ) and CAF lysates ( f , h ). qRT‒PCR analysis of MM2 and MMP12 mRNA expression in 344SQ cell ( d ) and CAF ( g ) samples. c–h 344SQ cells and CAFs were pretreated with an anti-integrin blocking antibody (anti-integrin αν, β3, or β5) before direct exposure to rWISP-1 (10 ng/ml for 344SQ cells, 3 ng/ml for CAFs) with or without TGF-β1 (10 ng/ml) for the indicated time. NS: not significant, ** P < 0.01, *** P < 0.001, two-tailed Student’s t test. The data are from one experiment representative of three independent experiments with similar results ( a , b left , c , e , f , h ) or from three independent experiments (mean ± standard error; a , b right , d , g )

Article Snippet: Gene overexpression experiments were performed with CAFs seeded at 0.5 × 10 6 cells/ml in 60 mm dishes for 24 h. For WISP-1 overexpression, cells were transfected with 2.5 μg of the mouse WISP-1 gene ORF cRNA clone expression plasmid (MG5A0255-CH, Sino Biological, North Wales, PA, USA) using Lipofectamine 2000 (Thermo Fisher Scientific, Waltham, MA, USA) for 24 h according to the manufacturer’s instructions.

Techniques: Migration, Microscopy, Blocking Assay, Western Blot, Expressing, Two Tailed Test

Journal: Cellular and Molecular Immunology

Article Title: Reprogramming of cancer-associated fibroblasts by apoptotic cancer cells inhibits lung metastasis via Notch1-WISP-1 signaling

doi: 10.1038/s41423-022-00930-w

Figure Lengend Snippet: Apoptotic lung cancer cells trigger Notch1 signaling for WISP-1 production in CAFs via enhanced Dll1 expression. a Immunoblot analysis of proteins in CAFs stimulated with or without ApoSQ for 20 h. b ELISA of WISP-1 in CAF CM, ApoSQ-CAF CM, and NecSQ-CAF CM. c , d qRT‒PCR analysis of Hes1 and WISP-1 in CAFs at the indicated time points after stimulation with ApoSQ or NecSQ. e 4×CSL luciferase assay in CAFs at the indicated time points after stimulation with ApoSQ or NecSQ. f Immunofluorescence staining for NICD1 (red) and WISP-1 (green; left ) and quantification ( right ) in CAFs stimulated with ApoSQ or NecSQ for 20 h. The imaging medium was VECTASHIELD fluorescent mounting medium containing DAPI. Original magnification: ×100. Scale bars = 100 μm. g Immunoblot analysis of Notch1 in CAFs transfected with control or Notch1 siRNA ( upper ). Densitometric analysis of the relative Notch1 abundance ( lower ). h Immunoblot analysis of the indicated proteins in CAFs transfected with Notch1 siRNA before stimulation with ApoSQ or NecSQ for 20 h. i ELISA of WISP-1 in CM from CAFs transfected with Notch1 siRNA before stimulation with or without ApoSQ for 20 h. Quantification of migrated and invaded 344SQ cells ( j ) and CAFs ( k ). j CAFs were transfected with control or Notch1 siRNA before exposure to ApoSQ for 20 h. CM was added to 344SQ cells with TGF-β1 (10 ng/ml) for 48 h. k CAFs were transfected with control or Notch1 siRNA before exposure to ApoSQ for 20 h, which the medium was then replaced with fresh medium containing TGF-β1 (10 ng/ml) for 24 h. l Flow cytometric analysis of Dll1, Dll3, Dll4, Jag1, and Jag2 expression in viable and UV-irradiated apoptotic cancer cells, including 344SQ, A549, and HCT116 cells. Mean fluorescence intensity (MFI) values ( left ) and histograms showing isotype control (gray) vs. specific Notch ligand antibody (red, blue, and green; right ) staining. m Immunoblot analysis of Notch ligand protein expresison in viable and UV-irradiated apoptotic cancer cells. n Immunoblot analysis of proteins in CAFs exposed to ApoSQ for 20 h in the presence or absence of a neutralizing anti-Dll1 antibody (20 μg/ml). o ELISA of WISP-1 in CM from CAFs exposed to ApoSQ for 20 h in the presence or absence of a neutralizing anti-Dll1 antibody. p Immunoblot analysis of Dll1 in 344SQ cells transfected with Dll1 siRNA ( upper ). Densitometric analysis of the relative Dll1 abundance ( lower ). q Immunoblot analysis of proteins in CAFs exposed to ApoSQ. r ELISA of WISP-1 in CM from CAFs exposed to ApoSQ. q , r 344SQ cells were transfected with control or Dll1 siRNA before apoptosis induction by UV irradiation. CAFs were then stimulated with or without ApoSQ for 20 h. NS: not significant, * P < 0.05, ** P < 0.01, *** P < 0.001, two-tailed Student’s t test. The data are from one experiment representative of three independent experiments with similar results ( a , f left , g upper , h , l right , m , n , p upper , q ) or from three independent experiments (mean ± standard error; b-e , f right , g lower , i – k , l left , o , p lower , r )

Article Snippet: Gene overexpression experiments were performed with CAFs seeded at 0.5 × 10 6 cells/ml in 60 mm dishes for 24 h. For WISP-1 overexpression, cells were transfected with 2.5 μg of the mouse WISP-1 gene ORF cRNA clone expression plasmid (MG5A0255-CH, Sino Biological, North Wales, PA, USA) using Lipofectamine 2000 (Thermo Fisher Scientific, Waltham, MA, USA) for 24 h according to the manufacturer’s instructions.

Techniques: Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Luciferase, Immunofluorescence, Staining, Imaging, Transfection, Irradiation, Fluorescence, Two Tailed Test

Journal: Cellular and Molecular Immunology

Article Title: Reprogramming of cancer-associated fibroblasts by apoptotic cancer cells inhibits lung metastasis via Notch1-WISP-1 signaling

doi: 10.1038/s41423-022-00930-w

Figure Lengend Snippet: BAI1/Rac1 signaling facilitates efferocytosis and participates in crosstalk with Notch1 signaling in CAFs. Phagocytic activity of CAFs toward ApoSQ in the absence or presence of annexin V (AnxV, 10 μg/ml; a ), transfected with BAI1 siRNA ( d ) or the BAI1-Flag construct ( g ), pretreated with a Rac1 inhibitor (NSC23766, 100 μM; k ), or transfected with Notch1 siRNA ( p ). Representative dot plots showing the percentages of phagocytosing CAFs (PKH26 + /PKH67 + ) and nonphagocytosing CAFs (PKH26 + /PKH67 - ) as indicated in the corresponding quadrants as determined by flow cytometry ( left ) and quantification of these populations ( right ). l Phagocytic activity of CAFs toward PKH67 + ApoSQ (green) in the absence or presence of NSC23766. The nucleus and actin cytoskeleton of CAFs were stained with DAPI (blue) and TRITC (tetramethylrhodamine)-conjugated phalloidin (red), respectively. Representative image of CAFs phagocytosing ApoSQ ( left ). The ROI − Merge panels show high-magnification images of regions of interest as indicated by the white squares in the Merge panels. Original magnification: ×200. Scale bars = 20 μm. The arrowheads indicate CAFs with engulfed apoptotic 344SQ cells or fragments. Phagocytosis was quantified by a phagocytic index based on confocal microscopy ( right ). Immunoblot analysis of the indicated proteins in CAFs exposed to ApoSQ for 20 h ( b , e , m ) or 2 h ( h ). c , f , i , n ELISA of WISP-1 in CAF CM and ApoSQ-CAF CM 20 h after exposure to ApoSQ. Relative Rac1 activity in CAFs at the indicated time points after ApoSQ stimulation ( j ) or transfection with Notch1 siRNA before ApoSQ stimulation for 24 h ( o ). NS: not significant, * P < 0.05, ** P < 0.01, *** P < 0.001, two-tailed Student’s t test. The data are from one experiment representative of three independent experiments with similar results ( a , d , g , k , l and p left , b , e , h , m ) or from three independent experiments (mean ± standard error; a , d , g , k , l and p right , c , f , i , j , n , o )

Article Snippet: Gene overexpression experiments were performed with CAFs seeded at 0.5 × 10 6 cells/ml in 60 mm dishes for 24 h. For WISP-1 overexpression, cells were transfected with 2.5 μg of the mouse WISP-1 gene ORF cRNA clone expression plasmid (MG5A0255-CH, Sino Biological, North Wales, PA, USA) using Lipofectamine 2000 (Thermo Fisher Scientific, Waltham, MA, USA) for 24 h according to the manufacturer’s instructions.

Techniques: Activity Assay, Transfection, Construct, Flow Cytometry, Staining, Confocal Microscopy, Western Blot, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: Cellular and Molecular Immunology

Article Title: Reprogramming of cancer-associated fibroblasts by apoptotic cancer cells inhibits lung metastasis via Notch1-WISP-1 signaling

doi: 10.1038/s41423-022-00930-w

Figure Lengend Snippet: Inhibition of Notch1 signaling reverses the increase in WISP-1 production and the inhibition of migration, invasion and signaling pathways in vivo. The experimental design was as described in Fig. . a – c Representative confocal images of primary tumor sections stained with an anti-α-SMA antibody (red; a ), an anti-NICD1 antibody (red; b ), and an anti-WISP-1 antibody (red; c ), an anti-Thy1 antibody (green), and DAPI (blue) ( left ). The ROI − Merge panels show high-magnification images of regions of interest as indicated by the white squares in the Merge panels. Original magnification: ×200. Scale bars = 20 μm. Quantification of α-SMA staining in Thy1 + cells and of NICD1 + cells and WISP-1 + cells among Thy1 + cells ( right ). The data are presented as the means ± standard errors from three mice per group ( a – c right ). d , e Representative confocal images of isolated Thy1 + CAFs from primary tumors stained with antibodies specific for α-SMA (red), NICD1 (red) or WISP-1 (green) ( left ). The imaging medium was VECTASHIELD fluorescent mounting medium containing DAPI. Images were acquired at ×200 magnification. Scale bars = 200 μm. Relative fluorescence intensity of α-SMA staining and quantification of NICD1 + cells and WISP-1 + cells ( right ). f ELISA of WISP-1 in the culture media of isolated Thy1 + CAFs, CD326 + tumor cells, and CD11b + TAMs and in serum. Phase contrast microscopy of migrated and invaded CD326 + tumor cells ( g ) and Thy1 + CAFs ( h ). Scale bars: 100 µm. i Immunoblot analysis of proteins in isolated CD326 + tumor cells. NS: not significant, * P < 0.05, ** P < 0.01, *** P < 0.001, Kruskal‒Wallis test with Dunn’s post hoc test. The data are from three replicates per condition, with cells pooled from three mice per replicate. The data are from one experiment representative of three independent experiments with similar results ( d and e left , g – i ) and are shown as the mean ± standard error from three independent experiments ( d and e right , f )

Article Snippet: Gene overexpression experiments were performed with CAFs seeded at 0.5 × 10 6 cells/ml in 60 mm dishes for 24 h. For WISP-1 overexpression, cells were transfected with 2.5 μg of the mouse WISP-1 gene ORF cRNA clone expression plasmid (MG5A0255-CH, Sino Biological, North Wales, PA, USA) using Lipofectamine 2000 (Thermo Fisher Scientific, Waltham, MA, USA) for 24 h according to the manufacturer’s instructions.

Techniques: Inhibition, Migration, In Vivo, Staining, Isolation, Imaging, Fluorescence, Enzyme-linked Immunosorbent Assay, Microscopy, Western Blot

Journal: Cellular and Molecular Immunology

Article Title: Reprogramming of cancer-associated fibroblasts by apoptotic cancer cells inhibits lung metastasis via Notch1-WISP-1 signaling

doi: 10.1038/s41423-022-00930-w

Figure Lengend Snippet: ApoSQ-CAF CM inhibits tumor growth and lung metastasis via WISP-1 in mice. a Schematic of the experimental design and group allocation. CAF CM, ApoSQ-CAF CM, ApoSQ-CAF CM + anti-WISP-1, or ApoSQ-CAF CM + IgG was intratumorally injected three times a week for 6 weeks starting 2 days after subcutaneous injection of 344SQ cells into syngeneic (129/Sν) mice ( n = 8 mice per group). Mice were necropsied 6 weeks later. Scatter plots of body weight ( b ), primary tumor weight ( d ), tumor volume ( e ), and numbers of lung metastatic nodules ( g ). c Representative images of primary tumors (yellow dashed circles). f Representative images of lungs with and without metastases. The yellow dashed circles indicate lung metastatic nodules. NS: not significant, * P < 0.05, ** P < 0.01, *** P < 0.001, Kruskal‒Wallis test with Dunn’s post hoc test. The data are presented as the mean ± standard error of results from 8 mice per group ( b , d , e , g ). h Numbers of mice with (w/) and without (w/o) visibly determined metastases (Met). i Heatmap showing differentially expressed tumor metastasis-related genes in CD326 + tumor cells isolated from primary tumors ( left ). Red: high expression; green: low expression. Relative expression of selected genes from PCR array profiling of tumor metastases ( right ). Log 2 fold change values (ApoSQ-CAF CM vs. CAF CM). j Heatmap showing differentially expressed genes encoding ECM and adhesion molecules in Thy1 + CAFs isolated from primary tumors ( left ). Relative expression levels of selected genes from PCR array profiling ( right ). Log 2 fold change values (ApoSQ-CAF CM vs. CAF CM). NS: not significant, * P < 0.05, ** P < 0.01, two-tailed Student’s t test. The data are from three replicates per condition with cells pooled from two or three mice per replicate (mean ± standard error; i and j right ). Phase contrast microscopy of migrated and invaded CD326 + tumor cells ( k ) and Thy1 + CAFs ( l ). Scale bars: 100 µ m. m Immunoblot analysis of proteins in CD326 + tumor cells. n qRT‒PCR analysis of CAF markers and MMPs in isolated Thy1 + CAFs. NS: not significant, * P < 0.05, ** P < 0.01, Kruskal‒Wallis test with Dunn’s post hoc test. The data are from three replicates per condition with cells pooled from two or three mice per replicate. The data are from one experiment representative of three independent experiments with similar results ( k–m ) or from three independent experiments (mean ± standard error; n )

Article Snippet: Gene overexpression experiments were performed with CAFs seeded at 0.5 × 10 6 cells/ml in 60 mm dishes for 24 h. For WISP-1 overexpression, cells were transfected with 2.5 μg of the mouse WISP-1 gene ORF cRNA clone expression plasmid (MG5A0255-CH, Sino Biological, North Wales, PA, USA) using Lipofectamine 2000 (Thermo Fisher Scientific, Waltham, MA, USA) for 24 h according to the manufacturer’s instructions.

Techniques: Injection, Isolation, Expressing, Two Tailed Test, Microscopy, Western Blot

Journal: Cellular and Molecular Immunology

Article Title: Reprogramming of cancer-associated fibroblasts by apoptotic cancer cells inhibits lung metastasis via Notch1-WISP-1 signaling

doi: 10.1038/s41423-022-00930-w

Figure Lengend Snippet: Recombinant WISP-1 inhibits tumor growth and lung metastasis in mice. a Schematic of the experimental design. Where indicated, rWISP-1 (12.5 and 25 μg/kg) was intratumorally injected three times a week for 6 weeks starting 2 days after subcutaneous injection of 344SQ cells into syngeneic (129/Sν) mice ( n = 8 mice per group). Mice were necropsied 6 weeks later. Scatter plots of body weight ( b ), primary tumor weight ( d ), tumor volume ( e ), and numbers of lung metastatic nodules ( g ). c Representative images of primary tumors (yellow dashed circles). f Representative images of lungs with and without metastases. The yellow dashed circles indicate lung metastatic nodules. NS: not significant, * P < 0.05, ** P < 0.01, Kruskal‒Wallis test with Dunn’s post hoc test. The data are presented as the mean ± standard error of results from 8 mice per group ( b , d , e , g ). h Numbers of mice with (w/) and without (w/o) visibly determined metastases (Met). Phase contrast microscopy of migrated and invaded CD326 + tumor cells ( i ) and Thy1 + CAFs ( j ). Scale bars : 100 µm. k Immunoblot analysis of proteins in CD326 + tumor cells. l qRT ‒ PCR analysis of CAF markers and MMPs in isolated Thy1 + CAFs. NS: not significant, * P < 0.05, ** P < 0.01, Kruskal‒Wallis test with Dunn’s post hoc test. The data are from three replicates per condition with cells pooled from two or three mice per replicate. The data are from one experiment representative of three independent experiments with similar results ( i–k ) or from three independent experiments (mean ± standard error; l )

Article Snippet: Gene overexpression experiments were performed with CAFs seeded at 0.5 × 10 6 cells/ml in 60 mm dishes for 24 h. For WISP-1 overexpression, cells were transfected with 2.5 μg of the mouse WISP-1 gene ORF cRNA clone expression plasmid (MG5A0255-CH, Sino Biological, North Wales, PA, USA) using Lipofectamine 2000 (Thermo Fisher Scientific, Waltham, MA, USA) for 24 h according to the manufacturer’s instructions.

Techniques: Recombinant, Injection, Microscopy, Western Blot, Isolation

Journal: American Journal of Physiology - Endocrinology and Metabolism

Article Title: Skeletal inflammation and attenuation of Wnt signaling, Wnt ligand expression, and bone formation in atherosclerotic ApoE-null mice

doi: 10.1152/ajpendo.00501.2015

Figure Lengend Snippet: TaqMan reagents used for quantification of mRNA by qPCR

Article Snippet: Transcript levels were calculated by normalizing to the housekeeping gene mitochondrial ribosomal protein S2 using the ΔC T method ( 34 ). table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 NCBI Gene Name Gene Name TaqMan # Alox12 Arachidonate 12-lipoxygenase Mm00545833_m1 Alox15 Arachidonate 15-lipoxygenase Mm01250458_m1 Axin2 Axin2 Mm00443610_m1 Mrps2 Mitochondrial ribosomal protein S2 (chob) Mm01962382_s91 Cx43 Gap junction membrane channel protein alpha 1 Mm01962382_s53 Dkk1 Dickkopf homolog 1 Mm00438422_m1 IL-1ß Interleukin-1β Mm00434228_m1 IL-6 Interleukin-6 Mm00446190_m1 Nkd2 Naked cuticle 2 homolog Mm00472240_m1 Sost Sclerostin Mm00470479_m1 Sfrp4 Secreted frizzled-related protein 4 Mm00840104_m1 TNF Tumor necrosis factor-α Mm00443258_m1 Tnfrsf11b Tumor necrosis factor receptor superfamily, member 11b (osteoprotogerin, OPG) Mm00435452_m1 Wisp1 WNT1 inducible signaling pathway protein 1 Mm00457574_m1 Wnt4 Wingless-related MMTV integration site 4 Mm00437341_m1 Wnt5a Wingless-related MMTV integration site 5A Mm00437347_m1 Wnt6 Wingless-related MMTV integration site 6 Mm00437353_m1 Wnt10b Wingless-related MMTV integration site 10B Mm00442104_m1 Wnt16 Wingless-related MMTV integration site 16 Mm00446420_m1 Open in a separate window TaqMan reagents used for quantification of mRNA by qPCR Statistics.

Techniques: Membrane

Journal: American Journal of Physiology - Endocrinology and Metabolism

Article Title: Skeletal inflammation and attenuation of Wnt signaling, Wnt ligand expression, and bone formation in atherosclerotic ApoE-null mice

doi: 10.1152/ajpendo.00501.2015

Figure Lengend Snippet: Effect of HFD on expression of Wnt target genes, Wnt ligands, and Wnt antagonists

Article Snippet: Transcript levels were calculated by normalizing to the housekeeping gene mitochondrial ribosomal protein S2 using the ΔC T method ( 34 ). table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 NCBI Gene Name Gene Name TaqMan # Alox12 Arachidonate 12-lipoxygenase Mm00545833_m1 Alox15 Arachidonate 15-lipoxygenase Mm01250458_m1 Axin2 Axin2 Mm00443610_m1 Mrps2 Mitochondrial ribosomal protein S2 (chob) Mm01962382_s91 Cx43 Gap junction membrane channel protein alpha 1 Mm01962382_s53 Dkk1 Dickkopf homolog 1 Mm00438422_m1 IL-1ß Interleukin-1β Mm00434228_m1 IL-6 Interleukin-6 Mm00446190_m1 Nkd2 Naked cuticle 2 homolog Mm00472240_m1 Sost Sclerostin Mm00470479_m1 Sfrp4 Secreted frizzled-related protein 4 Mm00840104_m1 TNF Tumor necrosis factor-α Mm00443258_m1 Tnfrsf11b Tumor necrosis factor receptor superfamily, member 11b (osteoprotogerin, OPG) Mm00435452_m1 Wisp1 WNT1 inducible signaling pathway protein 1 Mm00457574_m1 Wnt4 Wingless-related MMTV integration site 4 Mm00437341_m1 Wnt5a Wingless-related MMTV integration site 5A Mm00437347_m1 Wnt6 Wingless-related MMTV integration site 6 Mm00437353_m1 Wnt10b Wingless-related MMTV integration site 10B Mm00442104_m1 Wnt16 Wingless-related MMTV integration site 16 Mm00446420_m1 Open in a separate window TaqMan reagents used for quantification of mRNA by qPCR Statistics.

Techniques: Expressing