Journal: Scientific Reports

Article Title: SPP1 + macrophages promote colorectal cancer progression by activating JAK2/STAT3 signaling pathway

doi: 10.1038/s41598-025-21420-9

Figure Lengend Snippet: In vitro assays to verify the relationship between SPP1 + TAMs and JAK/STAT3 signaling pathway. The group descriptions of Ctrl, Positive, and Negative are the same as in Fig. . The DMSO group added DMSO to the medium to culture MC38 cells as a control. WP1066 group added WP1066 in medium; In the Pos + WP14066 group, WP1066-treated MC38 cells were cultured in SPP1 + TAMs conditioned medium. ( A , B ) WB detection of JAK/STAT3 pathway protein expression levels in the Control, Negative, and Positive groups. Compared with the Control group, **** P < 0.0001, and compared with the negative group, #### P < 0.0001; ( C ) CCK-8 assay to detect MC38 cell viability in each group; ( D ) TUNEL staining to detect the apoptosis rate of MC38 cells in each group; ( E – G ) Immunofluorescence to detect the expression of EMT-related molecules (E-cadherin, N-cadherin, fibronectin) in each group; H – I ) WB detection of JAK/STAT3 pathway proteins in the DMSO, WP1066, and Pos + WP1066 groups, Compared with the DMSO group, **** P < 0.0001.

Article Snippet: The cells were divided into the following six groups: Control group, where MC38 cells were cultured in RPMI-1640 medium alone; Negative or Positive group, where MC38 cells were cultured in SPP1 − TAMs or SPP1 + TAMs conditioned medium, respectively; DMSO group and WP1066 group, DMSO or JAK/STAT3 pathway inhibitor WP1066 (10 ng/ml, HY-15312, MCE) were added to MC38 cell medium, respectively.

Techniques: In Vitro, Control, Cell Culture, Expressing, CCK-8 Assay, TUNEL Assay, Staining, Immunofluorescence

Journal: Scientific Reports

Article Title: SPP1 + macrophages promote colorectal cancer progression by activating JAK2/STAT3 signaling pathway

doi: 10.1038/s41598-025-21420-9

Figure Lengend Snippet: In vitro assays to verify the relationship between SPP1 + TAMs and JAK/STAT3 signaling pathway. The group descriptions of Ctrl, Positive, and Negative are the same as in Fig. . The DMSO group added DMSO to the medium to culture MC38 cells as a control. WP1066 group added WP1066 in medium; In the Pos + WP14066 group, WP1066-treated MC38 cells were cultured in SPP1 + TAMs conditioned medium. ( A , B ) WB detection of JAK/STAT3 pathway protein expression levels in the Control, Negative, and Positive groups. Compared with the Control group, **** P < 0.0001, and compared with the negative group, #### P < 0.0001; ( C ) CCK-8 assay to detect MC38 cell viability in each group; ( D ) TUNEL staining to detect the apoptosis rate of MC38 cells in each group; ( E – G ) Immunofluorescence to detect the expression of EMT-related molecules (E-cadherin, N-cadherin, fibronectin) in each group; H – I ) WB detection of JAK/STAT3 pathway proteins in the DMSO, WP1066, and Pos + WP1066 groups, Compared with the DMSO group, **** P < 0.0001.

Article Snippet: The cells were divided into the following six groups: Control group, where MC38 cells were cultured in RPMI-1640 medium alone; Negative or Positive group, where MC38 cells were cultured in SPP1 − TAMs or SPP1 + TAMs conditioned medium, respectively; DMSO group and WP1066 group, DMSO or JAK/STAT3 pathway inhibitor WP1066 (10 ng/ml, HY-15312, MCE) were added to MC38 cell medium, respectively.

Techniques: In Vitro, Control, Cell Culture, Expressing, CCK-8 Assay, TUNEL Assay, Staining, Immunofluorescence

Journal: Scientific reports

Article Title: Interleukin-7 enhances recruitment of MDSCs by regulating MCP-1 via JAK1/STAT3 signaling pathway in non-small cell lung cancer.

doi: 10.1038/s41598-025-01868-5

Figure Lengend Snippet: Fig. 2. IL-7/IL-7R upregulates the expression of MCP-1 and affect phosphorylated form of JAK1 and STAT3. (A–F) A549 and H1299 cells were treated with IL-7 and A-IL-7R plus IL-7 for 24 h, then MCP-1 protein and mRNA were detected by western blot and qRT-PCR. (G–L) A549 and H1299 cells were treated with IL-7 and A-IL-7R plus IL-7 for 30 min (pJAK1 and JAK1) and 24 h (pSTAT3 and STAT3), then pJAK1, JAK1, pSTAT3 and STAT3 protein were detected by western blot. The data are shown as the mean ± SEM from the at least three independent experiments; *P < 0.05,**P < 0.01 and ***P < 0.001.

Article Snippet: Human and mouse recombinant IL-7, MCP-1were purchased from Peprotech (Peprotech, Rocky Hill, NJ), antiIL-7 receptor antibody (A-IL-7R) were from Santa (Santa Cruz, CA), CCR2 inhibitor RS102895, JAK1 inhibitor Ruxolitinib, STAT3 inhibitor WP1066 were from Med Chem Express (Monmouth Junction, NJ, USA).

Techniques: Expressing, Western Blot, Quantitative RT-PCR

Journal: Scientific reports

Article Title: Interleukin-7 enhances recruitment of MDSCs by regulating MCP-1 via JAK1/STAT3 signaling pathway in non-small cell lung cancer.

doi: 10.1038/s41598-025-01868-5

Figure Lengend Snippet: Fig. 3. Treatment of JAKl inhibitor Ruxolitinib suppresses IL-7-mediated elevation of MCP-1 expression. (A–J) A549 and H1299 cells were treated with IL-7 and Ruxolitinib plus IL-7(cell preincubated with IL-7 for 24 h then incubated with Ruxolitinib of 5 μM for 1 h), then pJAK1, JAK1, pSTAT3, STAT3 and MCP-1 protein were detected by western blot and the level of MCP-1 mRNA was detected by qRT-PCR. The data are shown as the mean ± SEM from the at least three independent experiments; *P < 0.05,**P < 0.01 and ***P < 0.001.

Article Snippet: Human and mouse recombinant IL-7, MCP-1were purchased from Peprotech (Peprotech, Rocky Hill, NJ), antiIL-7 receptor antibody (A-IL-7R) were from Santa (Santa Cruz, CA), CCR2 inhibitor RS102895, JAK1 inhibitor Ruxolitinib, STAT3 inhibitor WP1066 were from Med Chem Express (Monmouth Junction, NJ, USA).

Techniques: Expressing, Incubation, Western Blot, Quantitative RT-PCR

Journal: Scientific reports

Article Title: Interleukin-7 enhances recruitment of MDSCs by regulating MCP-1 via JAK1/STAT3 signaling pathway in non-small cell lung cancer.

doi: 10.1038/s41598-025-01868-5

Figure Lengend Snippet: Fig. 4. Treatment of STAT3 inhibitor WP1066 suppresses IL-7-mediated elevation of MCP-1 expression. (A– H) A549 and H1299 cells were treated with IL-7 and WP1066 plus IL-7(cell preincubated with IL-7 for 24 h then incubated with WP1066 of 10 μM for 1 h), then pSTAT3, STAT3 and MCP-1 protein were detected by western blot and the level of MCP-1 mRNA was detected by qRT-PCR. The data are shown as the mean ± SEM from the at least three independent experiments; *P < 0.05,**P < 0.01 and ***P < 0.001.

Article Snippet: Human and mouse recombinant IL-7, MCP-1were purchased from Peprotech (Peprotech, Rocky Hill, NJ), antiIL-7 receptor antibody (A-IL-7R) were from Santa (Santa Cruz, CA), CCR2 inhibitor RS102895, JAK1 inhibitor Ruxolitinib, STAT3 inhibitor WP1066 were from Med Chem Express (Monmouth Junction, NJ, USA).

Techniques: Expressing, Incubation, Western Blot, Quantitative RT-PCR

Journal: Scientific reports

Article Title: Interleukin-7 enhances recruitment of MDSCs by regulating MCP-1 via JAK1/STAT3 signaling pathway in non-small cell lung cancer.

doi: 10.1038/s41598-025-01868-5

Figure Lengend Snippet: Fig. 5. IL-7 promotes lung cancer development in vivo. (A) Representative images of LLC tumors from different groups. (B) Tumor volume curve was calculated after injection every three days. (C) The levels of expression of MCP-1 mRNA in mice tumor tissue were detected by qRT-PCR. (D) Representative images of immunohistochemical (IHC) staining for MCP-1, STAT3, pSTAT3, JAK1 and pJAK1. (E,F) Representative images of HE and immunohistochemical (IHC) staining for Ki-67. The index of Ki-67 was counted using Image software. In mouse lung cancer tissue specimens, JAK1, pJAK1, STAT3 and MCP-1 are mainly located in the cytoplasm of tumor cells, pSTAT3 is mainly located in the cytoplasm and nucleus of tumor cells, and Ki-67 is mainly located in the nucleus of tumor cells (100 magnification. Scale bar 100 μm). The data are shown as the mean ± SEM; *P < 0.05,**P < 0.01 and ***P < 0.001.

Article Snippet: Human and mouse recombinant IL-7, MCP-1were purchased from Peprotech (Peprotech, Rocky Hill, NJ), antiIL-7 receptor antibody (A-IL-7R) were from Santa (Santa Cruz, CA), CCR2 inhibitor RS102895, JAK1 inhibitor Ruxolitinib, STAT3 inhibitor WP1066 were from Med Chem Express (Monmouth Junction, NJ, USA).

Techniques: In Vivo, Injection, Expressing, Quantitative RT-PCR, Immunohistochemical staining, Immunohistochemistry, Software