primary antibodies against fibroblast activation protein fap  (Cell Signaling Technology Inc)

Journal: bioRxiv

Article Title: Bidirectional Crosstalk Between Bladder Cancer Cells and Normal Fibroblasts Drives Phenotypic Reprogramming and Modulates Chemosensitivity

doi: 10.1101/2025.09.09.675061

Figure Lengend Snippet: (a–c) Flow-cytometry workflow and representative plots for (a) NF alone, (b) NF co-cultured with RT112, and (c) NF co-cultured with T24 for 48 h. Gates show total cells (FSC-A vs SSC-A), singlets (FSC-H vs FSC-A), and the HBSF population identified by Alexa Fluor 488 negative (pre-labelled cancer with CellTracker Green). The frequency of gated HBSF within each condition is indicated. Right-hand histograms display activation-marker staining in gated HBSF (αSMA/FAP; Alexa Fluor 647 channel). (d) Quantification of mean fluorescence intensity (MFI) for αSMA and FAP in HBSF monoculture versus direct co-culture with RT112 or T24 (48 h). Both co-cultures significantly increased fibroblast activation marker expression, with the strongest induction observed in T24– HBSF. Bars show mean ± SD from n = 3.

Article Snippet: For NF, primary antibodies against fibroblast activation protein (FAP) and α-smooth muscle actin (αSMA) (Cell Signaling Technology) were applied at 1:100 overnight at 4°C.

Techniques: Flow Cytometry, Cell Culture, Activation Assay, Marker, Staining, Fluorescence, Co-Culture Assay, Expressing

Journal: Molecular Cancer

Article Title: circNOX4 activates an inflammatory fibroblast niche to promote tumor growth and metastasis in NSCLC via FAP/IL-6 axis

doi: 10.1186/s12943-024-01957-5

Figure Lengend Snippet: Suppression of IL-6 effectively restrains the tumor progression in vivo. A Schematic diagram illustrating that A549 cells were subcutaneously co-injected with NFs stably transfected with vector or circNOX4 into nude mice, and IL-6 antibody was locally injected twice a week. B Images of nude mice and xenograft tumors of each group ( n = 8). C Tumor growth curves were shown in different treatment groups. D Tumor weights were measured in different treatment groups. E Representative images of IHC for FAP, CD31, N-cadherin, Vimentin, MMP2, and MMP9 in xenograft tumors. Scale bar = 100 μm. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: After blocking the membranes with 5% milk-TBST, they were incubated with appropriate dilutions of specific primary antibodies against FAP (ab207178, 1:1000, Abcam) and α-SMA (ab124964, 1:1000, Abcam).

Techniques: In Vivo, Injection, Stable Transfection, Transfection, Plasmid Preparation

Journal: Cell death & disease

Article Title: IGF2BP2-meidated m 6 A modification of CSF2 reprograms MSC to promote gastric cancer progression.

doi: 10.1038/s41419-023-06163-7

Figure Lengend Snippet: Fig. 2 CSF2 regulates the pro-tumor phenotype and function of MSCs. A In vitro tropism of MSCs towards HGC-27 cells assessed via a transwell system. Original magnification: ×100. Scale bar = 50 μm. B qRT-PCR analysis of FAP and α-SMA expression in MSCs and P-MSCs. C Western blot analyses of FAP and α-SMA expression in MSCs and P-MSCs. D Luminex assay to determine the inflammatory cytokine profile in the supernatants from MSCs and P-MSCs. E Cell viability assay for HGC-27 cells treated with the supernatants from MSCs and P-MSCs. F Colony formation assays for HGC-27 cells treated with the supernatants from MSCs and P-MSCs. G Transwell migration assay for HGC-27 cells treated with the supernatants from MSCs and P-MSCs. Original magnification: ×100. Scale bar = 50 μm. H CCK-8 assay for IC50 of 5-FU in HGC-27 cells treated with the supernatants from MSCs and P-MSCs. I Flow cytometry analyses of the apoptotic rate of HGC-27 cells exposed to 5-FU. *P < 0.05; **P < 0.01; ***P < 0.001.

Article Snippet: Primary antibodies against FAP (#66562), α-SMA (#19245), Notch1 (#3608), CSF2 (#56712), ubiquitin (#20326) (Cell Signaling Technology, USA), and IGF2BP2 (11601-1-AP, Proteintech, Wuhan, China) were applied to the membranes for incubation. β-actin (CW0096M, Cwbio, Beijing, China) was used as the loading control.

Techniques: In Vitro, Quantitative RT-PCR, Expressing, Western Blot, Luminex, Viability Assay, Transwell Migration Assay, CCK-8 Assay, Flow Cytometry

Journal: Cell death & disease

Article Title: IGF2BP2-meidated m 6 A modification of CSF2 reprograms MSC to promote gastric cancer progression.

doi: 10.1038/s41419-023-06163-7

Figure Lengend Snippet: Fig. 4 IGF2BP2 modulates CSF2 m6A modification to induce MSC reprogramming. A In vitro tropism of MSCs towards HGC-27 cells assessed via a transwell system. Original magnification: ×100. Scale bar = 50 μm. B, C qRT-PCR (B) and western blot (C) analyses of FAP and α- SMA expression in MSCs from different groups. D Luminex analyses of the inflammatory factor profile in the supernatants from MSCs. E, F The proliferation of HGC-27 cells following exposure to the supernatants from different MSCs was assessed by cell viability (E) and colony formation assays (F). G Transwell assay for the migration of HGC-27 cells upon treatment with the supernatants from different MSCs. Original magnification: ×100. Scale bar = 50 μm. H CCK-8 assay for IC50 of 5-FU in HGC-27 cells treated with the supernatants from different MSCs. I Flow cytometric analyses of the apoptotic rate of HGC-27 cells after exposure to 5-FU. *P < 0.05; **P < 0.01; ***P < 0.001.

Article Snippet: Primary antibodies against FAP (#66562), α-SMA (#19245), Notch1 (#3608), CSF2 (#56712), ubiquitin (#20326) (Cell Signaling Technology, USA), and IGF2BP2 (11601-1-AP, Proteintech, Wuhan, China) were applied to the membranes for incubation. β-actin (CW0096M, Cwbio, Beijing, China) was used as the loading control.

Techniques: In Vitro, Quantitative RT-PCR, Western Blot, Expressing, Luminex, Transwell Assay, Migration, CCK-8 Assay

Journal: Cell death & disease

Article Title: IGF2BP2-meidated m 6 A modification of CSF2 reprograms MSC to promote gastric cancer progression.

doi: 10.1038/s41419-023-06163-7

Figure Lengend Snippet: Fig. 6 IGF2BP2/CSF2/Notch1 axis regulates MSC reprogramming. A In vitro tropism of MSCs towards HGC-27 cells assessed via a transwell system. Original magnification: ×100. Scale bar = 50 μm. B, C qRT-PCR (B) and western blot (C) were performed to analyze the expression of FAP and α-SMA in different MSCs. D Luminex assay was employed to examine the inflammatory factor profile in MSC supernatant from each group. E, F Cell viability assays (E) and colony formation assays (F) were performed to assess the proliferation of HGC-27 cells treated with the supernatants from different MSCs. G Transwell assays were performed to evaluate the migration of HGC-27 cells treated with MSC supernatant from each group. Original magnification: ×100. Scale bar = 50 μm. H CCK-8 assay for IC50 of 5-FU in HGC-27 cells. *P < 0.05; **P < 0.01; ***P < 0.001.

Article Snippet: Primary antibodies against FAP (#66562), α-SMA (#19245), Notch1 (#3608), CSF2 (#56712), ubiquitin (#20326) (Cell Signaling Technology, USA), and IGF2BP2 (11601-1-AP, Proteintech, Wuhan, China) were applied to the membranes for incubation. β-actin (CW0096M, Cwbio, Beijing, China) was used as the loading control.

Techniques: In Vitro, Quantitative RT-PCR, Western Blot, Expressing, Luminex, Migration, CCK-8 Assay