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Osteosarcoma neutrophil elastase (ELA2) promotes osteosarcoma cell proliferation in vitro and in vivo . (A) Colony formation assay showed that ELA2 accelerated osteosarcoma proliferation in U2OS and 143B cells (unpaired two-tailed Student’s t -test, N = 3). (B) Immunofluorescence image showed that Lv-ELA2 was successfully transfected into <t>K7M2</t> cell lines and stably expressed (scale bar: 100 μm). (C) Quantitative analysis of relative ELA2 expression level in the Lv-Ctrl group and Lv-ELA2 group by qPCR assays (unpaired two-tailed Student’s t -test, N = 3). (D) The results of the cell cycle in the Ctrl group and ELA2-OE group using flow cytometry (left). The statistical analysis of G0/G1 phase, S phase, and G2/M phase cell proportions (right) (unpaired two-tailed Student’s t -test, N = 3). (E) Images of tumors from the Ctrl group and ELA2-OE group tumors ( N = 3 for each group). (F) Tumor weight and tumor volume were measured and quantitatively analyzed (tumor volume = length × width 2 /2; unpaired two-tailed Student’s t -test, N = 3). (G) Images of Ki-67 + immunohistochemistry staining in the Ctrl group and ELA2-OE group (scale bar: 100 μm). ( *** p < 0.001).
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Osteosarcoma neutrophil elastase (ELA2) promotes osteosarcoma cell proliferation in vitro and in vivo . (A) Colony formation assay showed that ELA2 accelerated osteosarcoma proliferation in U2OS and 143B cells (unpaired two-tailed Student’s t -test, N = 3). (B) Immunofluorescence image showed that Lv-ELA2 was successfully transfected into <t>K7M2</t> cell lines and stably expressed (scale bar: 100 μm). (C) Quantitative analysis of relative ELA2 expression level in the Lv-Ctrl group and Lv-ELA2 group by qPCR assays (unpaired two-tailed Student’s t -test, N = 3). (D) The results of the cell cycle in the Ctrl group and ELA2-OE group using flow cytometry (left). The statistical analysis of G0/G1 phase, S phase, and G2/M phase cell proportions (right) (unpaired two-tailed Student’s t -test, N = 3). (E) Images of tumors from the Ctrl group and ELA2-OE group tumors ( N = 3 for each group). (F) Tumor weight and tumor volume were measured and quantitatively analyzed (tumor volume = length × width 2 /2; unpaired two-tailed Student’s t -test, N = 3). (G) Images of Ki-67 + immunohistochemistry staining in the Ctrl group and ELA2-OE group (scale bar: 100 μm). ( *** p < 0.001).
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wt cells  (ATCC)
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Osteosarcoma neutrophil elastase (ELA2) promotes osteosarcoma cell proliferation in vitro and in vivo . (A) Colony formation assay showed that ELA2 accelerated osteosarcoma proliferation in U2OS and 143B cells (unpaired two-tailed Student’s t -test, N = 3). (B) Immunofluorescence image showed that Lv-ELA2 was successfully transfected into <t>K7M2</t> cell lines and stably expressed (scale bar: 100 μm). (C) Quantitative analysis of relative ELA2 expression level in the Lv-Ctrl group and Lv-ELA2 group by qPCR assays (unpaired two-tailed Student’s t -test, N = 3). (D) The results of the cell cycle in the Ctrl group and ELA2-OE group using flow cytometry (left). The statistical analysis of G0/G1 phase, S phase, and G2/M phase cell proportions (right) (unpaired two-tailed Student’s t -test, N = 3). (E) Images of tumors from the Ctrl group and ELA2-OE group tumors ( N = 3 for each group). (F) Tumor weight and tumor volume were measured and quantitatively analyzed (tumor volume = length × width 2 /2; unpaired two-tailed Student’s t -test, N = 3). (G) Images of Ki-67 + immunohistochemistry staining in the Ctrl group and ELA2-OE group (scale bar: 100 μm). ( *** p < 0.001).
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Osteosarcoma neutrophil elastase (ELA2) promotes osteosarcoma cell proliferation in vitro and in vivo . (A) Colony formation assay showed that ELA2 accelerated osteosarcoma proliferation in U2OS and 143B cells (unpaired two-tailed Student’s t -test, N = 3). (B) Immunofluorescence image showed that Lv-ELA2 was successfully transfected into <t>K7M2</t> cell lines and stably expressed (scale bar: 100 μm). (C) Quantitative analysis of relative ELA2 expression level in the Lv-Ctrl group and Lv-ELA2 group by qPCR assays (unpaired two-tailed Student’s t -test, N = 3). (D) The results of the cell cycle in the Ctrl group and ELA2-OE group using flow cytometry (left). The statistical analysis of G0/G1 phase, S phase, and G2/M phase cell proportions (right) (unpaired two-tailed Student’s t -test, N = 3). (E) Images of tumors from the Ctrl group and ELA2-OE group tumors ( N = 3 for each group). (F) Tumor weight and tumor volume were measured and quantitatively analyzed (tumor volume = length × width 2 /2; unpaired two-tailed Student’s t -test, N = 3). (G) Images of Ki-67 + immunohistochemistry staining in the Ctrl group and ELA2-OE group (scale bar: 100 μm). ( *** p < 0.001).
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Osteosarcoma neutrophil elastase (ELA2) promotes osteosarcoma cell proliferation in vitro and in vivo . (A) Colony formation assay showed that ELA2 accelerated osteosarcoma proliferation in U2OS and 143B cells (unpaired two-tailed Student’s t -test, N = 3). (B) Immunofluorescence image showed that Lv-ELA2 was successfully transfected into K7M2 cell lines and stably expressed (scale bar: 100 μm). (C) Quantitative analysis of relative ELA2 expression level in the Lv-Ctrl group and Lv-ELA2 group by qPCR assays (unpaired two-tailed Student’s t -test, N = 3). (D) The results of the cell cycle in the Ctrl group and ELA2-OE group using flow cytometry (left). The statistical analysis of G0/G1 phase, S phase, and G2/M phase cell proportions (right) (unpaired two-tailed Student’s t -test, N = 3). (E) Images of tumors from the Ctrl group and ELA2-OE group tumors ( N = 3 for each group). (F) Tumor weight and tumor volume were measured and quantitatively analyzed (tumor volume = length × width 2 /2; unpaired two-tailed Student’s t -test, N = 3). (G) Images of Ki-67 + immunohistochemistry staining in the Ctrl group and ELA2-OE group (scale bar: 100 μm). ( *** p < 0.001).

Journal: Frontiers in Immunology

Article Title: Neutrophil elastase promotes low molecular weight cyclin E1 formation to accelerate osteosarcoma proliferation

doi: 10.3389/fimmu.2025.1647913

Figure Lengend Snippet: Osteosarcoma neutrophil elastase (ELA2) promotes osteosarcoma cell proliferation in vitro and in vivo . (A) Colony formation assay showed that ELA2 accelerated osteosarcoma proliferation in U2OS and 143B cells (unpaired two-tailed Student’s t -test, N = 3). (B) Immunofluorescence image showed that Lv-ELA2 was successfully transfected into K7M2 cell lines and stably expressed (scale bar: 100 μm). (C) Quantitative analysis of relative ELA2 expression level in the Lv-Ctrl group and Lv-ELA2 group by qPCR assays (unpaired two-tailed Student’s t -test, N = 3). (D) The results of the cell cycle in the Ctrl group and ELA2-OE group using flow cytometry (left). The statistical analysis of G0/G1 phase, S phase, and G2/M phase cell proportions (right) (unpaired two-tailed Student’s t -test, N = 3). (E) Images of tumors from the Ctrl group and ELA2-OE group tumors ( N = 3 for each group). (F) Tumor weight and tumor volume were measured and quantitatively analyzed (tumor volume = length × width 2 /2; unpaired two-tailed Student’s t -test, N = 3). (G) Images of Ki-67 + immunohistochemistry staining in the Ctrl group and ELA2-OE group (scale bar: 100 μm). ( *** p < 0.001).

Article Snippet: 143B (CVCL_2270), KHOS (CVCL_2546), HOS (CVCL_0312), MG63 (CVCL_0426), U2OS (CVCL_0042), and SaoS2 (CVCL_0548) cells were human osteosarcoma cell lines, and K7M2 cells were mouse osteosarcoma cell lines; all were purchased from the American Type Culture Collection.

Techniques: In Vitro, In Vivo, Colony Assay, Two Tailed Test, Immunofluorescence, Transfection, Stable Transfection, Expressing, Flow Cytometry, Immunohistochemistry, Staining