Journal: Biochemistry and Biophysics Reports

Article Title: Hepatoma-derived growth factor and non-coding RNA network in ovarian cancer patients

doi: 10.1016/j.bbrep.2025.102168

Figure Lengend Snippet: ROC of CA125, HE4 and, HDGF.

Article Snippet: Enzyme-linked immunosorbent assays (ELISAs) were performed to measure serum levels of HDGF (BT Laboratories, China), HE4 (Xema, Russia), and CA125 (Pishtaz-Teb, Iran) using commercially available kits, following the manufacturers’ protocols.

Techniques:

Journal: Nature Cell Biology

Article Title: Peritumoural adipose tissue drives immune evasion in colorectal cancer via adipose–mesenchymal transformation

doi: 10.1038/s41556-026-01885-0

Figure Lengend Snippet: a . Representative images of MC38-OVA (left) and tumour growth (right) in C57BL/6J mice ( n = 5). b . Representative images of CT26 tumours (left) and tumour growth (right) in BALB/c mice ( n = 5). c . Schematic diagram illustrating the working principle of ATS-GNP adipocytolysis. ATS-GNP, containing a CaCO 3 core, is receptor-mediated for endocytosis. Upon reaching the acidic environment, ATS-GNP releases CO 2 gas, disrupting the adipose cell membrane. d . Tumour growth of MC38 tumours in C57BL/6J, BALB/c-Nude (left), and NSG (right) mice ( n = 5). e . Representative images of E0771 tumours at day 16 of the experiment in C57BL/6J mice following removal of PAT ( n = 5). f . Tumour weights of E0771 tumours at day 16 in C57BL/6J mice with the removal of PAT ( n = 5). g . Representative flow cytometry plots of CD45 + cells gated on live cells in MC38-OVA tumours. h . Violin plots showing the expression of CXCR7 across all cell types in CRC patients, analysed using the Kruskal-Wallis test. i . Editing strategy for constructing Cxcl12 fl/fl cKO mouse. j . Experimental design for constructing Control and Cxcl12 fl/fl cKO mice bearing MC38-OVA tumours near PAT. k . Western blot analysis for verifying the knockout efficiency of Cxcl12 in PAT of Control mice and Cxcl12 fl/fl cKO mice. l . RT-qPCR analysis for verifying the Cxcl12 knockout efficiency in liver, spleen, uterus, and tumour tissues from Control mice and Cxcl12 fl/fl cKO mice ( n = 6). Data represent ≥ 3 independent experiments. P -values were calculated using two-way ANOVA with Tukey’s correction for multiple comparisons ( a -right, b -right, and d ), two-way ANOVA with Bonferroni’s correction for multiple comparisons ( f ), or a two-sided, unpaired Student’s t -test ( l ). Graphs display mean ± SD ( a , b , d , f , l ). Panels created with BioRender : c , i and j , Huaiqiang, J. https://biorender.com/cy7rgjm (2026).

Article Snippet: For drug treatment, a CXCL12 neutralizing antibody (Merck Millipore) and an anti-mouse PD-1 antibody (Bio X Cell) were administered via intraperitoneal injection.

Techniques: Membrane, Flow Cytometry, Expressing, Control, Western Blot, Knock-Out, Quantitative RT-PCR

Journal: Nature Cell Biology

Article Title: Peritumoural adipose tissue drives immune evasion in colorectal cancer via adipose–mesenchymal transformation

doi: 10.1038/s41556-026-01885-0

Figure Lengend Snippet: a , Cell–cell communication analysis based on ligand–receptor interactions (top six) between stromal cells and lymphocytes in tVAT (left) and a comparison between tVAT, dVAT and tumour (right). b , Marked CXCL12–CXCR4 interactions among CD8 + T cells, CD4 + T cells, B cells, plasma cells and stromal cell populations in tVAT, dVAT and tumour. The width of the lines represents the probability of communication. c , Violin plots showing the expression of CXCL12 (top) and CXCR4 (bottom) across all cell types in patients with CRC. d , Violin plots comparing the expression of CXCL12 in dVAT versus tVAT (top) and tumour versus tVAT (bottom) in patients with CRC, analysed using a two-sided Wilcoxon test. e , Experimental design for the PAT C57BL/6J mouse model treated with IgG or anti-CXCL12 antibody (left), and representative MC38 tumour images at day 16 of the experiment (right) ( n = 5). f , Tumour growth (left) and tumour weights (right) of MC38 tumours at day 16 of the experiment in C57BL/6J mice ( n = 5). g , Representative MC38 tumour images (left) and tumour growth (right) of experiments in control and Cxcl12 fl/fl cKO mice ( n = 6). h , Tumour weights of MC38 tumours in control and Cxcl12 fl/fl cKO mice at day 16 of the experiment ( n = 6). i , Flow cytometry analysis of the infiltration of various CXCR4 + immune cells in MC38 tumours in Control and Cxcl12 fl/fl cKO mice ( n = 6). j , Schematic diagram of the chemotaxis assay using T cells as ‘sensors’ and conditional medium as a ‘sink’ (left), and the aggregated trajectories of control or CXCL12-induced T cells migrating for 1 h (right). k , Quantitative analysis of CXCR4 + CD45.1 + T cells in MC38 tumours with and without removal of PAT or contralateral inguinal adipose tissue (control) by flow cytometry ( n = 5). Data represent ≥3 independent experiments. Statistical significance was assessed by a two-sided permutation test ( a ), two-sided unpaired Student’s t -test ( f right, h and i ), one-way analysis of variance (ANOVA) with Tukey’s correction for multiple comparisons ( k right) or two-way ANOVA ( f left and g right). Graphs display mean ± s.d. ( f – i , k ). Panels created with BioRender : e and k , Huaiqiang, J. https://biorender.com/ovq2e39 (2026).

Article Snippet: For drug treatment, a CXCL12 neutralizing antibody (Merck Millipore) and an anti-mouse PD-1 antibody (Bio X Cell) were administered via intraperitoneal injection.

Techniques: Comparison, Clinical Proteomics, Expressing, Control, Flow Cytometry, Chemotaxis Assay

Journal: Nature Cell Biology

Article Title: Peritumoural adipose tissue drives immune evasion in colorectal cancer via adipose–mesenchymal transformation

doi: 10.1038/s41556-026-01885-0

Figure Lengend Snippet: a . UMAP plot of VAT-associated stromal cells in tVAT and dVAT from CRC patients. The 8 clusters, labelled by inferred cell types, are denoted by colour. b . Dot plot showing RNA expression of marker genes used to define pAC, adCAF, and APC subclusters. Circle size represents the log-normalized P -value, while colour intensity indicates the log-transformed mean expression of marker genes. c . Sorting strategy for isolating adCAFs from mouse PAT by flow cytometry. d . RNA sequencing analysis of adCAF markers in sorted adCAF-enriched stromal cells and non-adCAF stromal cells ( n = 4). Statistical significance was assessed using a two-sided, unpaired Student’s t -test. e . RNA sequencing analysis of adipogenesis markers in sorted adCAF-enriched stromal cells and non-adCAF stromal cells ( n = 4). Statistical significance was assessed using a two-sided, unpaired Student’s t -test. f . Representative images of the morphology of sorted adCAF-enriched stromal cells and non-adCAF stromal cells. Scale bar = 500 μm. g . Cross-tissue interactions analysis based on ligand-receptor pairs between adCAFs and tumour cells (left) and a comparison between tVAT and dVAT (right). A two-sided permutation test was used to determine the significance of pathways. h . Western blot analysis of CXCL12 protein expression in sorted adCAF-enriched stromal cells and non-adCAF stromal cells ( n = 3). The data are presented as a box-and-whisker graph (bounds of box: first to third quartile, bottom and top line: minimum to maximum, central line: median) for ( d - e ).

Article Snippet: For drug treatment, a CXCL12 neutralizing antibody (Merck Millipore) and an anti-mouse PD-1 antibody (Bio X Cell) were administered via intraperitoneal injection.

Techniques: RNA Expression, Marker, Transformation Assay, Expressing, Flow Cytometry, RNA Sequencing, Comparison, Western Blot, Whisker Assay

Journal: Nature Cell Biology

Article Title: Peritumoural adipose tissue drives immune evasion in colorectal cancer via adipose–mesenchymal transformation

doi: 10.1038/s41556-026-01885-0

Figure Lengend Snippet: a , UMAP of all stromal cells in tVAT, dVAT, tumour and normal from patients with CRC, with ten clusters labelled by inferred cell types. Major lineages included ASCs, pACs, CAFs, pericytes (PCs) and mesothelial cells (Mesos). b , Heatmap displaying the distribution of eight stromal cell subtypes across different tissue types. c , UMAP of eight subsets of VAT-associated stromal cells in tVAT and dVAT from patients with CRC, including ASCs, pACs and adCAFs. d , Beeswarm plot showing the distribution and abundance of VAT-associated stromal cell types in Nhoods between tVAT and dVAT. e , Stack plot displaying the abundance of the eight VAT-associated stromal cell subsets in dVAT and tVAT. f , Heatmap showing the RNA expression of various marker genes in VAT-associated stromal cell types, including ASC/pAC markers, CAF markers, cytokines and stromal markers. g , Representative multiplex immunofluorescence images showing the presence of adCAFs in tVAT samples from patients with CRC. Scale bar, 10 μm. DAPI, 4,6-diamidino-2-phenylindole. h , Relative expression levels of multiple cytokines and protumoural factors in adCAF-enriched stromal cells ( n = 4) and non-adCAF stromal cells ( n = 4) derived from the PAT of mice xenograft models. The data are presented as a box-and-whisker graph (bounds of box show first to third quartile, bottom and top line show minimum to maximum and the central line shows the median). i , Cell–cell communication analysis based on ligand–receptor pairs (top six) between adCAFs and lymphocytes in tVAT (left) and a comparison between tVAT and dVAT (right). j , RT–qPCR (left) and ELISA (right) detecting the RNA expression and protein secretion of CXCL12 in sorted adCAF-enriched stromal cells and non-adCAF stromal cells from PAT of mice. Data represent ≥3 independent experiments. All data are shown as mean ± s.d. and statistical significance was assessed by a two-sided, unpaired Wilcoxon test ( h ), two-sided permutation test ( i ) and Student’s t -test ( j ).

Article Snippet: For drug treatment, a CXCL12 neutralizing antibody (Merck Millipore) and an anti-mouse PD-1 antibody (Bio X Cell) were administered via intraperitoneal injection.

Techniques: RNA Expression, Marker, Multiplex Assay, Immunofluorescence, Expressing, Derivative Assay, Whisker Assay, Comparison, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: Nature Cell Biology

Article Title: Peritumoural adipose tissue drives immune evasion in colorectal cancer via adipose–mesenchymal transformation

doi: 10.1038/s41556-026-01885-0

Figure Lengend Snippet: a . UMAP of all cells in tVAT and dVAT from 5 CRC patients, with 16 clusters labelled by inferred cell types. Major lineages included various immunocytes, VAT-associated stromal cells, adipocytes (ACs), endothelial cells (ECs), pericytes (PCs) and mesothelial cells (Mesos). b . Heatmap of representative marker genes across all cell populations. c . UMAP of 6 subsets of ACs in tVAT and dVAT from 5 CRC patients. d . Beeswarm plot showing the distribution and abundance of ACs in Nhoods between tVAT and dVAT. e . Box plot to compare the abundance of ACs in dVAT ( n = 5) and tVAT ( n = 5). P -values were calculated using a two-side Wilcoxon test. f . Cell-cell communication analysis based on ligand-receptor interactions (top 3) between CD8 + T cells and adipocytes, and between CD8 + T cells and VAT-associated stromal cells in tVAT. A two-sided permutation test was used to determine the significance of pathways. Only significant ligand-receptor pairs ( P < 0.05) are shown. Dot size encodes the interaction score, and colour scale indicates the communication probability. g . Violin plots showing the expression of CXCL12, VAT-associated markers, and CAF-associated markers in adipocytes and VAT-associated stromal cells from CRC patients. h . UMAP of 8 subsets of VAT-associated stromal cells in tVAT and dVAT from 5 CRC patients, including APCs, pACs and adipocyte-derived cancer-associated fibroasts (adCAFs). The 8 clusters, labelled by inferred cell types, are denoted by colour. i . Beeswarm plot showing the distribution and abundance of VAT-associated stromal cell types in Nhoods between tVAT ( n = 5) and dVAT ( n = 5). j . Box plot to compare the abundance of VAT-associated stromal cells in dVAT and tVAT. P -values were calculated using a two-side Wilcoxon test. k . Heatmap showing the RNA expression patterns of representative marker genes across all cell populations, including CAF markers, stromal markers, APC/pAC markers, white/brown/beige fat markers, and cytokines and growth markers. The data are presented as a box-and-whisker graph (bounds of box: first to third quartile, bottom and top line: minimum to maximum, central line: median) for ( e , j ).

Article Snippet: For drug treatment, a CXCL12 neutralizing antibody (Merck Millipore) and an anti-mouse PD-1 antibody (Bio X Cell) were administered via intraperitoneal injection.

Techniques: Marker, Expressing, Derivative Assay, RNA Expression, Whisker Assay

Journal: Nature Cell Biology

Article Title: Peritumoural adipose tissue drives immune evasion in colorectal cancer via adipose–mesenchymal transformation

doi: 10.1038/s41556-026-01885-0

Figure Lengend Snippet: a , Experimental design for constructing Control and Mdk DTR cKO mice bearing MC38 tumours near PAT, followed by αPD-1 therapy. b , c , Representative MC38 tumour images ( b ), tumour weights ( c left) and tumour growth ( c right) in control and Mdk DTR cKO mice treated with IgG or anti-PD-1 ( n = 6). d , Flow cytometry analysis of the infiltration of immunocytes, including T cells, CD4 + T cells, CD8 + T cells, and tumour-specific CD8 + T cells, CXCR4 + immunocytes, CXCR4 + T cell, CXCR4 + CD4 + T cell, CXCR4 + CD8 + T cell and CXCR4 + tumour-specific T cell in MC38-OVA tumours from the four treatment groups ( n = 6). e , f , Representative MC38 tumour images ( e ) and tumour weights ( f ) of the experiment in mice treated with anti-CXCL12 and/or anti-PD-1 ( n = 5). g , Representative MRI image of CRC tumour and corresponding tVAT area region of CR and non-CR patients pre- and post-immuno-chemoradiotherapy. The yellow area represents the tVAT area, whereas the red area denotes the tumour region. Note that the mass visible in the intestinal lumen (top right) is faecal material. h , Pre-treatment tVAT area difference based on 3D Slicer between CR ( n = 30) and non-CR ( n = 37) patients. The data are presented as a box-and-whisker graph (bounds of box show first to third quartile, bottom and top line show minimum to maximum and the central line shows the median). i , ROC plot of response predicting ability of pre-treatment PAT area in immuno-chemoradiotherapy of proficient mismatch repair patients with CRC, compared with conventional indexes, including CPS, TPS, CEA and CA199 ( n = 67) with optimal cutoff. j , Comparison of pCR ratio in tVAT high and low group according to the optimal cutoff. k , Graphical abstract depicting how tumours reshape the stromal environment in tVAT and how tVAT competes for immunocytes from the tumour to promote immune escape. Data represent ≥3 independent experiments. Statistical significance was assessed using a two-sided, unpaired Student’s t -test ( d , h ), one-way ANOVA with Tukey’s correction for multiple comparisons ( c left, f ) or two-way ANOVA with Tukey’s correction for multiple comparisons ( c right). Graphs display mean ± s.d. ( c , d , f , h ). Panels created with BioRender : a and k , Huaiqiang, J. https://biorender.com/e5jwcye (2026).

Article Snippet: For drug treatment, a CXCL12 neutralizing antibody (Merck Millipore) and an anti-mouse PD-1 antibody (Bio X Cell) were administered via intraperitoneal injection.

Techniques: Control, Flow Cytometry, Whisker Assay, Comparison

Journal: Nature Cell Biology

Article Title: Peritumoural adipose tissue drives immune evasion in colorectal cancer via adipose–mesenchymal transformation

doi: 10.1038/s41556-026-01885-0

Figure Lengend Snippet: a . Editing strategy for constructing Mdk DTR mouse. b . Flow cytometry representative plots and bar graphs for verifying the elimination efficiency of adCAFs (FAP + PDGFRB + MDK + stromal cells) in PAT of Control mice and Mdk DTR cKO mice. c . Experimental design for combination therapy with αCXCL12 and αPD-1 in mice bearing MC38-OVA tumours near PAT. d . MC38 tumour weights of the experiment in mice treated with αCXCL12 and/or αPD-1 ( n = 6). e . Flow cytometry analysis of the infiltration of immunocytes, including T cells, CD4 + T cells, CD8 + T cells, and tumour-specific CD8 + T cells in MC38-OVA tumours from the 4 treatment groups ( n = 6). f . Flow cytometry analysis of the infiltration of CXCR4 + immunocytes, including CXCR4 + T cell, CXCR4 + CD4 + T cell and CXCR4 + CD8 + T cell in MC38-OVA tumours from the 4 treatment groups ( n = 6). g . Boxplot of difference of tVAT area in CR and non-CR patients with T3 or T4 stage separately. The data are presented as a box-and-whisker graph (bounds of box: first to third quartile, bottom and top line: minimum to maximum, central line: median). h . ROC plot of prediction ability of tVAT area in patients with T3 or T4 stage separately. Data represent ≥ 3 independent experiments. P -values were calculated using a two-sided, unpaired Student’s t -test ( b , e - g ) and two-way ANOVA with Tukey’s correction for multiple comparisons ( d ). Graphs display mean ± SD ( b , d , e - g ). Panels created with BioRender : a and c , Huaiqiang, J. https://biorender.com/33r2gmm (2026).

Article Snippet: For drug treatment, a CXCL12 neutralizing antibody (Merck Millipore) and an anti-mouse PD-1 antibody (Bio X Cell) were administered via intraperitoneal injection.

Techniques: Flow Cytometry, Control, Whisker Assay

Journal: Nature Cell Biology

Article Title: Peritumoural adipose tissue drives immune evasion in colorectal cancer via adipose–mesenchymal transformation

doi: 10.1038/s41556-026-01885-0

Figure Lengend Snippet: a . Experimental design for combination therapy with AMD3100 and αPD-1 in mice bearing MC38-OVA tumours near subcutaneous PAT. b . Representative MC38 tumour images of the mice treated with AMD3100 and/or αPD-1 ( n = 5). c . Representative MC38 tumour weights of the mice treated with AMD3100 and/or αPD-1 ( n = 5). d . Representative MC38 tumour growth of the mice treated with AMD3100 and/or αPD-1 ( n = 5). e . Representative MC38 tumour images in MC38-OVA caecal orthotopic tumour-bearing mice treated with AMD3100 and/or αPD-1 ( n = 5). f . Representative MC38 tumour weights in MC38-OVA caecal orthotopic tumour-bearing mice treated with AMD3100 and/or αPD-1 ( n = 5). g . Representative MC38 Bioluminescence Images in MC38-OVA caecal orthotopic tumour-bearing mice treated with AMD3100 and/or αPD-1 ( n = 5). h . Representative MC38 tumour images in MC38-OVA caecal orthotopic tumour-bearing mice treated with αCXCL12 and/or αPD-1 ( n = 5). i . Representative MC38 tumour weights in MC38-OVA caecal orthotopic tumour-bearing mice treated with αCXCL12 and/or αPD-1 ( n = 5). j . Representative MC38 Bioluminescence Images in MC38-OVA caecal orthotopic tumour-bearing mice treated with αCXCL12 and/or αPD-1 ( n = 5). k . Flow cytometry analysis of the infiltration of immunocytes, including T cells, CD4 + T cells, CD8 + T cells, and tumour-specific CD8 + T cells in MC38-OVA tumours from 4 treatment groups ( n = 5). l . Flow cytometry analysis of the infiltration of CXCR4 + immunocytes, including CXCR4 + T cell, CXCR4 + CD4 + T cell and CXCR4 + CD8 + T cell in MC38-OVA tumours from 4 treatment groups ( n = 5). Data represent ≥ 3 independent experiments. P -values were calculated using a two-sided, unpaired Student’s t -test ( c , f , i , k , l ) and two-way ANOVA with Tukey’s correction for multiple comparisons ( d ). Graphs display mean ± SD ( c - d , f , i , k - l ). Panel created with BioRender : a , Huaiqiang, J. https://biorender.com/wbcolts (2026).

Article Snippet: For drug treatment, a CXCL12 neutralizing antibody (Merck Millipore) and an anti-mouse PD-1 antibody (Bio X Cell) were administered via intraperitoneal injection.

Techniques: Flow Cytometry

Journal: BMC Molecular and Cell Biology

Article Title: EphB4/ TNFR2/ERK/MAPK signaling pathway comprises a signaling axis to mediate the positive effect of TNF-α on osteogenic differentiation

doi: 10.1186/s12860-020-00273-2

Figure Lengend Snippet: Primer sequences for qRT-PCR

Article Snippet: The anti-mouse primary antibodies used in this study were listed as following: RUNX2 (1:1000, catalog no. 12556S; CST, Danvers, MA, USA), BSP (1:1000, catalog no. 5468S CST, Danvers, MA,USA), EphB4 (1:1000, catalog no. A00690; Boster, China), TNFR2 (1:1000, catalog no. ab19139; abcam, Danvers, MA, USA), p38 (1:1000,catalog no. ab170099; abcam, Danvers, MA, USA), p -p38 (1:1000,catalog no. ab195049; abcam, Danvers, MA, USA), JNK1 + 2 + 3 (1:1000,catalog no. ab179461; abcam, Danvers, MA, USA), p -JNK1 + 2 + 3 (1:5000,catalog no. ab124956; abcam, Danvers, MA, USA), ERK1/2 (1:10000,catalog no. ab184699; abcam, Danvers, MA, USA), and p -ERK1/2 (1:8000,catalog no. ab76299; abcam, Danvers, MA, USA).

Techniques:

Journal: BMC Molecular and Cell Biology

Article Title: EphB4/ TNFR2/ERK/MAPK signaling pathway comprises a signaling axis to mediate the positive effect of TNF-α on osteogenic differentiation

doi: 10.1186/s12860-020-00273-2

Figure Lengend Snippet: The effect of the lentivirus-mediated shRNA interference of TNFR2 on TNF-α-stimulated EphB4 expression and osteogenic differentiation. a MC3T3-E1 cells stably transduced with lentiviral particles were selected with puromycin and named as pHBLV-TNFR2siRNA1 cells, pHBLV-TNFR2siRNA2 cells, pHBLV-TNFR2siRNA3 cells and pHBLV-NC cells, respectively. The mRNA levels of Tnfr2 were determined in these cells, among which the pHBLV-TNFR2siRNA1 cells displayed the highest TNFR2 gene silencing efficiency and were selected to continue the following studies. b TNFR2 protein levels in pHBLV-TNFR2siRNA1 cells and pHBLV-NC cells. c , d mRNA levels of Ephb4 , Runx2 and Bsp in pHBLV-TNFR2siRNA1 cells and pHBLV-NC cells cultured in the osteogenic induction medium supplemented with 0.5 ng/ml TNF-α for 24 h ( c ) or 48 h ( d ). e , f Protein levels of EphB4, RUNX2 and BSP in pHBLV-TNFR2siRNA1 cells and pHBLV-NC cells cultured in the osteogenic induction medium supplemented with 0.5 ng/ml TNF-α for 24 h ( e ) or 48 h ( f ). *, p < 0.05 vs. the pHBLV-NC group; **, p < 0.01 vs. the pHBLV-NC group

Article Snippet: The anti-mouse primary antibodies used in this study were listed as following: RUNX2 (1:1000, catalog no. 12556S; CST, Danvers, MA, USA), BSP (1:1000, catalog no. 5468S CST, Danvers, MA,USA), EphB4 (1:1000, catalog no. A00690; Boster, China), TNFR2 (1:1000, catalog no. ab19139; abcam, Danvers, MA, USA), p38 (1:1000,catalog no. ab170099; abcam, Danvers, MA, USA), p -p38 (1:1000,catalog no. ab195049; abcam, Danvers, MA, USA), JNK1 + 2 + 3 (1:1000,catalog no. ab179461; abcam, Danvers, MA, USA), p -JNK1 + 2 + 3 (1:5000,catalog no. ab124956; abcam, Danvers, MA, USA), ERK1/2 (1:10000,catalog no. ab184699; abcam, Danvers, MA, USA), and p -ERK1/2 (1:8000,catalog no. ab76299; abcam, Danvers, MA, USA).

Techniques: shRNA, Expressing, Stable Transfection, Transduction, Cell Culture

Journal: BMC Molecular and Cell Biology

Article Title: EphB4/ TNFR2/ERK/MAPK signaling pathway comprises a signaling axis to mediate the positive effect of TNF-α on osteogenic differentiation

doi: 10.1186/s12860-020-00273-2

Figure Lengend Snippet: The effect of the impaired binding between TNF-α and TNFR2 on TNF-α-stimulated EphB4 expression and osteogenic differentiation. MC3T3-E1 cells were treated with an anti-mouse TNFR2/ CD120b/TNFRSF1B neutralizing antibody (TNFR2 NAb) at the concentration of 0.2 μg/ml, and were cultured in the osteogenic induction medium supplemented with or without 0.5 ng/ml TNF-α. Cells treated with 0.2 μg/ml of the normal rabbit IgG negative control antibody (control Ab) served as negative controls. (a) ALP activities were determined 7d or 14d after the treatment. (b, c) mRNA levels of Ephb4 , Runx2 and Bsp were determined after 24 h (b) or 48 h (c). (d, e) Protein levels of EphB4, RUNX2 and BSP were determined after 24 h (d) or 48 h (e). a, p < 0.05 vs. the control Ab group; b, p < 0.05 vs. the TNFR2 NAb group; c, p < 0.05 vs. the TNF-α + control Ab group

Article Snippet: The anti-mouse primary antibodies used in this study were listed as following: RUNX2 (1:1000, catalog no. 12556S; CST, Danvers, MA, USA), BSP (1:1000, catalog no. 5468S CST, Danvers, MA,USA), EphB4 (1:1000, catalog no. A00690; Boster, China), TNFR2 (1:1000, catalog no. ab19139; abcam, Danvers, MA, USA), p38 (1:1000,catalog no. ab170099; abcam, Danvers, MA, USA), p -p38 (1:1000,catalog no. ab195049; abcam, Danvers, MA, USA), JNK1 + 2 + 3 (1:1000,catalog no. ab179461; abcam, Danvers, MA, USA), p -JNK1 + 2 + 3 (1:5000,catalog no. ab124956; abcam, Danvers, MA, USA), ERK1/2 (1:10000,catalog no. ab184699; abcam, Danvers, MA, USA), and p -ERK1/2 (1:8000,catalog no. ab76299; abcam, Danvers, MA, USA).

Techniques: Binding Assay, Expressing, Concentration Assay, Cell Culture, Negative Control

Journal: BMC Molecular and Cell Biology

Article Title: EphB4/ TNFR2/ERK/MAPK signaling pathway comprises a signaling axis to mediate the positive effect of TNF-α on osteogenic differentiation

doi: 10.1186/s12860-020-00273-2

Figure Lengend Snippet: The effect of inhibited EphB4 forward signaling on TNF-α-stimulated TNFR2 expression and osteogenic differentiation. (a) A potent inhibitor of EphB4 auto-phosphorylation, NVP-BHG712, was used to suppress EphB4 forward signaling. MC3T3-E1 cells were pretreated with 200 nM NVP-BHG712 in the regular culture medium for 1 h. Cells were then incubated in osteogenic induction medium supplemented with 200 nM NVP-BHG712 and/or 0.5 ng/ml TNF-α for 7d or 14d. MC3T3-E1 cells cultured in osteogenic induction medium served as controls. The ALP activities were determined. (b, c) MC3T3-E1 cells were pretreated with 200 nM NVP-BHG712 for 1 h in the regular culture medium, and then incubated in osteogenic induction medium supplemented with 200 nM NVP-BHG712 and/or 0.5 ng/ml TNF-α. Cells cultured in osteogenic induction medium served as controls. mRNA levels of Tnfr2 , Runx2 and Bsp were determined after 24 h (b) or 48 h (c) of incubation. (d, e) MC3T3-E1 cells were pretreated with 200 nM NVP-BHG712 for 1 h in the regular culture medium, and then incubated in osteogenic induction medium supplemented with 200 nM NVP-BHG712 and/or 0.5 ng/ml TNF-α. Cells cultured in osteogenic induction medium served as controls. Protein levels of TNFR2, RUNX2 and BSP were determined after 24 h (d) or 48 h (e) of incubation. a, p < 0.05 vs. the control group; b, p < 0.05 vs. the NVP-BHG712 group; c, p < 0.05 vs. the TNF-α group

Article Snippet: The anti-mouse primary antibodies used in this study were listed as following: RUNX2 (1:1000, catalog no. 12556S; CST, Danvers, MA, USA), BSP (1:1000, catalog no. 5468S CST, Danvers, MA,USA), EphB4 (1:1000, catalog no. A00690; Boster, China), TNFR2 (1:1000, catalog no. ab19139; abcam, Danvers, MA, USA), p38 (1:1000,catalog no. ab170099; abcam, Danvers, MA, USA), p -p38 (1:1000,catalog no. ab195049; abcam, Danvers, MA, USA), JNK1 + 2 + 3 (1:1000,catalog no. ab179461; abcam, Danvers, MA, USA), p -JNK1 + 2 + 3 (1:5000,catalog no. ab124956; abcam, Danvers, MA, USA), ERK1/2 (1:10000,catalog no. ab184699; abcam, Danvers, MA, USA), and p -ERK1/2 (1:8000,catalog no. ab76299; abcam, Danvers, MA, USA).

Techniques: Expressing, Incubation, Cell Culture

Journal: BMC Molecular and Cell Biology

Article Title: EphB4/ TNFR2/ERK/MAPK signaling pathway comprises a signaling axis to mediate the positive effect of TNF-α on osteogenic differentiation

doi: 10.1186/s12860-020-00273-2

Figure Lengend Snippet: EphB4, TNFR2 and MAPK signaling pathways comprise a signaling axis to mediate the positive effect of TNF-α on osteogenic differentiation. a Levels of p38, p -p38, ERK1/2, p -ERK1/2, JNK1 + 2 + 3 and p -JNK1 + 2 + 3 in MC3T3-E1 cells treated with TNF-α for 0 min, 5 min, 15 min, 30 min and 60 min. b Levels of p38, p -p38, ERK1/2, p -ERK1/2, JNK1 + 2 + 3 and p -JNK1 + 2 + 3 in the pHBLV-TNFR2siRNA1 cells and the pHBLV-NC cells treated with or without 0.5 ng/ml TNF-α in regular culture medium for 15 min. c MC3T3-E1 cells were pretreated with or without 200 nM NVP-BHG712 in the regular culture medium for 1 h, and then 0.5 ng/ml TNF-α was added into the medium. The cells were incubated for another 15 min. Levels of ERK1/2 and p -ERK1/2 were determined. d-f MC3T3-E1 cells were cultured in the regular culture medium and pretreated with the ERK inhibitor U0126 (10 μM) for 1 h. The culture medium was then switched to the osteogenic induction medium supplemented with 0.5 ng/ml TNF-α and U0126 (10 μM). Cells treated without U0126 (10 μM) served as controls. ALP activities were determined 7d or 14d after the treatment ( d ). mRNA levels ( e ) and protein levels ( f ) of BSP and RUNX2 were determined 3 days after the treatment. *, p < 0.05 vs. the control group; **, p < 0.01 vs. the control group

Article Snippet: The anti-mouse primary antibodies used in this study were listed as following: RUNX2 (1:1000, catalog no. 12556S; CST, Danvers, MA, USA), BSP (1:1000, catalog no. 5468S CST, Danvers, MA,USA), EphB4 (1:1000, catalog no. A00690; Boster, China), TNFR2 (1:1000, catalog no. ab19139; abcam, Danvers, MA, USA), p38 (1:1000,catalog no. ab170099; abcam, Danvers, MA, USA), p -p38 (1:1000,catalog no. ab195049; abcam, Danvers, MA, USA), JNK1 + 2 + 3 (1:1000,catalog no. ab179461; abcam, Danvers, MA, USA), p -JNK1 + 2 + 3 (1:5000,catalog no. ab124956; abcam, Danvers, MA, USA), ERK1/2 (1:10000,catalog no. ab184699; abcam, Danvers, MA, USA), and p -ERK1/2 (1:8000,catalog no. ab76299; abcam, Danvers, MA, USA).

Techniques: Incubation, Cell Culture

Journal: BMC Molecular and Cell Biology

Article Title: EphB4/ TNFR2/ERK/MAPK signaling pathway comprises a signaling axis to mediate the positive effect of TNF-α on osteogenic differentiation

doi: 10.1186/s12860-020-00273-2

Figure Lengend Snippet: Schematic diagram of the EphB4, TNFR2 and ERK/MAPK signaling pathways. A low concentration of TNF-α first enhances the expression of EphB4, which in turn promoted the expression of TNFR2. The elevated TNFR2 level leads to the activation of the ERK signaling pathway, which eventually enhances the osteogenic differentiation of MC3T3-E1 cells. TNFR2, tumor necrosis factor receptor 2; TNF-α, tumor necrosis factor-alpha; MAPK, mitogen-activated protein kinase; ERK, extracellular signal regulated kinase

Article Snippet: The anti-mouse primary antibodies used in this study were listed as following: RUNX2 (1:1000, catalog no. 12556S; CST, Danvers, MA, USA), BSP (1:1000, catalog no. 5468S CST, Danvers, MA,USA), EphB4 (1:1000, catalog no. A00690; Boster, China), TNFR2 (1:1000, catalog no. ab19139; abcam, Danvers, MA, USA), p38 (1:1000,catalog no. ab170099; abcam, Danvers, MA, USA), p -p38 (1:1000,catalog no. ab195049; abcam, Danvers, MA, USA), JNK1 + 2 + 3 (1:1000,catalog no. ab179461; abcam, Danvers, MA, USA), p -JNK1 + 2 + 3 (1:5000,catalog no. ab124956; abcam, Danvers, MA, USA), ERK1/2 (1:10000,catalog no. ab184699; abcam, Danvers, MA, USA), and p -ERK1/2 (1:8000,catalog no. ab76299; abcam, Danvers, MA, USA).

Techniques: Concentration Assay, Expressing, Activation Assay

Journal: BMC Cancer

Article Title: Cancer-associated fibroblasts induce sorafenib resistance of hepatocellular carcinoma cells through CXCL12/FOLR1

doi: 10.1186/s12885-023-11613-8

Figure Lengend Snippet: CAFs induce sorafenib resistance in HCC cells by secreting CXCL12. a The results of immunofluorescence showed that the expression of CXCL12 in CAFs in HCC tissues was significantly higher than that in paracancerous tissues (left). Statistical plot of fluorescence intensity of fibroblasts expressing α-SMA and CXCL12 in HCC tissues and paracancerous tissues (right). b ELISA showed that CAFs secreted higher level of CXCL12 than NFs. c , d Colony forming assays detected the sorafenib resistance of HCC cells (HepG2 and Huh7), after treated with the cellular supernatant of CAFs and NFs, sorafenib, and AMD3100. e , f Flow cytometry apoptosis assay detected the sorafenib resistance of HCC cells (HepG2 and Huh7), after treated with the cellular supernatant of CAFs and NFs, sorafenib, and AMD3100. g , h Western blotting was performed to detect the expression of β-actin, and Cleaved Caspase-3 in HCC cells (HepG2 and Huh7), which were treated with the cellular supernatant of CAFs, sorafenib, and AMD3100. The data presented mean ± SEM. * p < 0.01; ** p < 0.001; *** p < 0.0001; **** p < 0.00001

Article Snippet: Primary antibodies were used for IHC staining: CXCL12 (Boster, BA1389, 1:100), and α-SMA (Abcam, ab119952, 1:100).

Techniques: Immunofluorescence, Expressing, Fluorescence, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Apoptosis Assay, Western Blot

Journal: BMC Cancer

Article Title: Cancer-associated fibroblasts induce sorafenib resistance of hepatocellular carcinoma cells through CXCL12/FOLR1

doi: 10.1186/s12885-023-11613-8

Figure Lengend Snippet: CXCL12 induces sorafenib resistance in HCC cells by up-regulating the expression of FOLR1. a We found two datasets of cancer cells treated with CXCL12 protein (GSE15893 and GSE40017) in the GEO database. We took the intersection of the differentially expressed genes between these two datasets and combined them with the reported drug-resistant genes of HCC to obtain two genes. FOLR1 was the most significantly upregulated drug resistance-related gene upon CXCL12 treatment. b The qPCR was performed to detect the level of FOLR1 in Huh7 and HepG2, which treated with CXCL12 protein and AMD3100. c Western blotting was performed to detect the expression of β-actin and CXCR4 in CXCR4 knockdown HCC cells (Huh7 and HepG2). d , e Western blotting was performed to detect the expression of β-actin, CXCR4, FOLR1, and Cleaved Caspase-3 in Huh7 and HepG2, after treated with sorafenib, CXCL12 protein, and AMD3100. f , g Western blotting was performed to detect the expression of β-actin, CXCR4, FOLR1, and Cleaved Caspase-3 in Huh7 and HepG2, after treated with sorafenib, AMD3100, the supernatant of CAFs, and NFs. h , i Colony forming assay detected the sorafenib resistance of HCC cells (HepG2 and Huh7), after treated with sorafenib, CXCL12 protein, the supernatant of CAFs, and AMD3100. j , k Western blotting was performed to detect the expression of β-actin, FOLR1, CXCR4, and Cleaved Caspase-3 in Huh7 and HepG2, after treated with sorafenib, anti-CXCR4, CXCL12 protein, and the supernatant of CAFs. The data presented mean ± SEM. ** p < 0.001; *** p < 0.0001; **** p < 0.00001

Article Snippet: Primary antibodies were used for IHC staining: CXCL12 (Boster, BA1389, 1:100), and α-SMA (Abcam, ab119952, 1:100).

Techniques: Expressing, Western Blot, Knockdown

Journal: BMC Cancer

Article Title: Cancer-associated fibroblasts induce sorafenib resistance of hepatocellular carcinoma cells through CXCL12/FOLR1

doi: 10.1186/s12885-023-11613-8

Figure Lengend Snippet: CAFs enhance sorafenib resistance of HCC cells through CXCL12 in vivo. a Representative images of tumors in mice of CAFs + AMD3100 group, CAFs group, and NFs group after different treatments. b The tumor volume in different treatment groups. c The tumor proliferation trend in different treatment groups. d Pathological validation of tumors under a microscope (40X), after H&E staining and Immunohistochemistry in tumor tissues. The immunohistochemistry staining to detect the expression of Cleaved Caspase-3 in different treatment groups from the tumor tissues of mice. e The expression level of Cleaved Caspase-3 in different treatment groups from the tumors of mice. The data presented mean ± SEM. ** p < 0.001; *** p < 0.0001; **** p < 0.00001

Article Snippet: Primary antibodies were used for IHC staining: CXCL12 (Boster, BA1389, 1:100), and α-SMA (Abcam, ab119952, 1:100).

Techniques: In Vivo, Biomarker Discovery, Microscopy, Staining, Immunohistochemistry, Expressing