Journal: Frontiers in Medicine

Article Title: Detection of Gas6/AXL complex and its expression changes in patients with ST-segment elevation myocardial infarction

doi: 10.3389/fmed.2025.1653708

Figure Lengend Snippet: (A) Binding of different antibodies to AXL. Binding of different antibodies to AXL and their interaction with Gas6. (B) List of full length and different truncations of AXL. (C) αAXL-9# and αAXL-13# were analyzed via flow cytometry for reactivity towards hAXL or hAXL truncations. (D) αAXL-4# were analyzed via flow cytometry for reactivity towards hAXL or hAXL truncations with or without Gas6-his.

Article Snippet: Generally, plates were coated with αAXL-4# antibody or AF154 (DY154, commercial kit from R&D system) or Gas6 antibody (DY885B, commercial kit from R&D Systems) overnight at 4°C, blocked with PBS containing 0.05% Tween and 0.1% BSA for 1 h at room temperature, and then incubated with plasma or standard protein (AXL-Gas6 complex, AXL protein or Gas6 protein from commercial kit) for 2 h at room temperature.

Techniques: Binding Assay, Flow Cytometry

Journal: Frontiers in Medicine

Article Title: Detection of Gas6/AXL complex and its expression changes in patients with ST-segment elevation myocardial infarction

doi: 10.3389/fmed.2025.1653708

Figure Lengend Snippet: The binding characteristic of AXL-4#. (A) Representation of interplay of AXL antibody and Gas6. (B) 293T-hAXL cells were incubated with isotype control (left), AXL-4# (middle) or AXL4# with Gas6 (right), then stained with PE-goat anti mice IgG2a. (C) 293T-memGas6 cells were incubated with anti-Gas6l (left), AXL-4# (middle) or AXL4# with Gas6 (right), then stained with PE-goat anti mice IgG2a. (D,E) The structure of αAXL-4# recognizing Gas6/AXL complex predicted by AlphaFold.

Article Snippet: Generally, plates were coated with αAXL-4# antibody or AF154 (DY154, commercial kit from R&D system) or Gas6 antibody (DY885B, commercial kit from R&D Systems) overnight at 4°C, blocked with PBS containing 0.05% Tween and 0.1% BSA for 1 h at room temperature, and then incubated with plasma or standard protein (AXL-Gas6 complex, AXL protein or Gas6 protein from commercial kit) for 2 h at room temperature.

Techniques: Binding Assay, Incubation, Control, Staining

Journal: Frontiers in Medicine

Article Title: Detection of Gas6/AXL complex and its expression changes in patients with ST-segment elevation myocardial infarction

doi: 10.3389/fmed.2025.1653708

Figure Lengend Snippet: Application of αAXL-4# in flow cytometry. (A,B) The binding of αAXL-4# to Gas6/AXL complex of different Species. (C,D) asDCs and other cell lines were incubated with αAXL-9#, αAXL-13 and αAXL-4#.

Article Snippet: Generally, plates were coated with αAXL-4# antibody or AF154 (DY154, commercial kit from R&D system) or Gas6 antibody (DY885B, commercial kit from R&D Systems) overnight at 4°C, blocked with PBS containing 0.05% Tween and 0.1% BSA for 1 h at room temperature, and then incubated with plasma or standard protein (AXL-Gas6 complex, AXL protein or Gas6 protein from commercial kit) for 2 h at room temperature.

Techniques: Flow Cytometry, Binding Assay, Incubation

Journal: Frontiers in Medicine

Article Title: Detection of Gas6/AXL complex and its expression changes in patients with ST-segment elevation myocardial infarction

doi: 10.3389/fmed.2025.1653708

Figure Lengend Snippet: Application of αAXL-4# in ELISA. (A) Different concentrations of AXL (0-1 μg/mL) with a single dose of Gas6 (0.25 μg/mL, red column) or without Gas6 (empty column) were added subsequently. (B) Different concentrations of Gas6 (0–1 μg/mL) with a single dose of AXL (1 μg/mL, red column) or without AXL (empty column) were added subsequently.

Article Snippet: Generally, plates were coated with αAXL-4# antibody or AF154 (DY154, commercial kit from R&D system) or Gas6 antibody (DY885B, commercial kit from R&D Systems) overnight at 4°C, blocked with PBS containing 0.05% Tween and 0.1% BSA for 1 h at room temperature, and then incubated with plasma or standard protein (AXL-Gas6 complex, AXL protein or Gas6 protein from commercial kit) for 2 h at room temperature.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Medicine

Article Title: Detection of Gas6/AXL complex and its expression changes in patients with ST-segment elevation myocardial infarction

doi: 10.3389/fmed.2025.1653708

Figure Lengend Snippet: Different levels of AXL-Gas6, AXL and Gas6 in human plasma. (A–C) Plates were coated by AXL-4# (A) , commercial AXL antibody (B) or commercial Gas6 antibody (C) respectively. The levels of AXL-Gas6, AXL, or Gas6 in human plasma were detected by ELISA.

Article Snippet: Generally, plates were coated with αAXL-4# antibody or AF154 (DY154, commercial kit from R&D system) or Gas6 antibody (DY885B, commercial kit from R&D Systems) overnight at 4°C, blocked with PBS containing 0.05% Tween and 0.1% BSA for 1 h at room temperature, and then incubated with plasma or standard protein (AXL-Gas6 complex, AXL protein or Gas6 protein from commercial kit) for 2 h at room temperature.

Techniques: Clinical Proteomics, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Medicine

Article Title: Detection of Gas6/AXL complex and its expression changes in patients with ST-segment elevation myocardial infarction

doi: 10.3389/fmed.2025.1653708

Figure Lengend Snippet: ROC curve and association with AXL/AXL-Gas6 with clinical data. (A) ROC curve; (B) AXL was associated with Cr in STEMI; (C–G) Gas6/AXL complex was associated with PLT, WBC, HDL, CK-MB and SO-FMC as indicated.

Article Snippet: Generally, plates were coated with αAXL-4# antibody or AF154 (DY154, commercial kit from R&D system) or Gas6 antibody (DY885B, commercial kit from R&D Systems) overnight at 4°C, blocked with PBS containing 0.05% Tween and 0.1% BSA for 1 h at room temperature, and then incubated with plasma or standard protein (AXL-Gas6 complex, AXL protein or Gas6 protein from commercial kit) for 2 h at room temperature.

Techniques:

Journal: bioRxiv

Article Title: AXL-GAS6/PROS1 Interaction: A Critical Switch Between Aberrant- and Healthy Repair Following Alveolar Lung Injury

doi: 10.1101/2025.04.07.647567

Figure Lengend Snippet: (A) IPF signaling pathway was one of the increasingly upregulated pathways over time as revealed by IPA analysis. The gene set contributing to this pathway included distinct groups of early and late profibrotic associated genes (marked by black rectangles) that were serially and time dependently upregulated. Timepoint order forced, genes hierarchical clustered. (B- D) Fold change of mRNA expression levels of TAM RTK family, Axl , Mertk and Tyro3 over the course of repair. (E-F) Fold change of expression level of TAM RTK ligands, Gas6 and Pros1 over the course of repair. Data are shown as mean ± SEM of n= 3-7. P ≤ 0.05 (*); P ≤ 0.01 (**); P ≤ 0.001 (***); P ≤ 0.0001 (****) compared with stuffer control.

Article Snippet: GAS6 was measured in human BALF or SAEC supernatant using the human GAS6 DuoSet ELISA kit (R&D Systems, #DY885B) and the human PROS1 ELISA Kit (Aviva Systems Biology, #OKBB010207).

Techniques: Expressing, Control

Journal: bioRxiv

Article Title: AXL-GAS6/PROS1 Interaction: A Critical Switch Between Aberrant- and Healthy Repair Following Alveolar Lung Injury

doi: 10.1101/2025.04.07.647567

Figure Lengend Snippet: (A) Schematic of the e xperimental setup to study AXL-GAS6/PROS1 interaction using aFRET in mouse lung tissue sections. Lungs were isolated on day 4 post DT. aFRET is a coincidence assay, labels both receptor (AXL) and ligands (GAS6 or PROS1) (see Methods for details). FRET occurs in the range of 1-10 nm. Scheme was created in BioRender. Geillinger-kästle, K. (2025) https://BioRender.com/h45x771 (B) Representative image of AXL-GAS6 interaction. (C) Representative image of AXL-PROS1 interaction. (B-C) Upper figures showed the mean of the donor lifetime (Ƭ) in the absence of acceptor. Lower figures showed the interaction state between AXL-GAS6/ PROS1 for each coincidental region (marked by yellow and black circles). Different coincidental regions possessed different interaction state. (D) Box and whiskers plots showed the heterogeneity of AXL-GAS6/PROS1 interaction states represented as FRET efficiency (Ef%). Ef% below 4% represents the Förster Radius (R 0 at 5.83nm) and not proteins, interactive state as marked by red dashed line. Data are shown as box and whiskers plot of n=5-6. Each dot represents median of Ef% for each mouse. P ≤ 0.05 (*). (E) Correlation plot between Ef% AXL-GAS6 and AXL expression. (F) Correlation plot between Ef% AXL-PROS1 and AXL expression. (G-H) Surface plasmon resonance response curve showing the binding of rhGAS6 and rhPROS1 in different concentration to rhAXL. (I-J) Concentration dependent binding of rhGAS6 and rhPROS1 to rhAXL. Different colored lines in (G-H) correspond to different colored dots in (I-J), which indicate different concentrations of rhGAS6 and rhPROS1. (K-L) GAS6 and PROS1 concentration in human BALF of non- diseased control, ILD/IPF-, and COPD patients. Data are shown as mean ± SEM of n= 4-8. P ≤ 0.01 (**); P ≤ 0.001 (***).

Article Snippet: GAS6 was measured in human BALF or SAEC supernatant using the human GAS6 DuoSet ELISA kit (R&D Systems, #DY885B) and the human PROS1 ELISA Kit (Aviva Systems Biology, #OKBB010207).

Techniques: Isolation, Expressing, SPR Assay, Binding Assay, Concentration Assay, Control

Journal: bioRxiv

Article Title: AXL-GAS6/PROS1 Interaction: A Critical Switch Between Aberrant- and Healthy Repair Following Alveolar Lung Injury

doi: 10.1101/2025.04.07.647567

Figure Lengend Snippet: (A) 7664 is a tissue section from a healthy lung whereas 1015 and 1033 are sections from IPF patients with various advanced degrees of IPF. The panels show representative images mapping the interactive states of GAS6 and PROS1 with AXL (pseudo-colour heat map of (Ef%) on the expression level of GAS6 and PROS1 (fluorescence images in grey scale). The individual violin plots quantify the heterogeneity of interactive states for both ligands and the receptor. The dotted line at 4% Ef represents the Förster Radius (R 0 ). Values below 4% are not representative of ligand-receptor interactions. (B) Global violin plots of all coincident regions (per pixel) of AXL-GAS6 and AXL-PROS1. Each global violin plot represents 2x10 6 data points. To determine the p values for AXL-GAS6 and AXL-PROS1, using non-parametric Mann- Whitney U test, we utilised 1000 randomly generated data points of the 2x10 6 . Healthy lung has similar distribution of AXL-GAS6 and AXL-PROS1 interactions, whereas the two patient samples show, in one case (1015) no interaction of AXL-GAS6 or AXL-PROS1 and another case (1033) a significantly higher distribution of AXL-PROS1 versus AXL-GAS6. The p values between AXL-GAS6 and AXL-PROS1 are 1.9x10 -3 (7664), 2.4x10 -1 (1015) and 8.2x10 - respectively. The dotted line at 4% Ef represents the Förster Radius (R 0 ).

Article Snippet: GAS6 was measured in human BALF or SAEC supernatant using the human GAS6 DuoSet ELISA kit (R&D Systems, #DY885B) and the human PROS1 ELISA Kit (Aviva Systems Biology, #OKBB010207).

Techniques: Expressing, Fluorescence, MANN-WHITNEY, Generated

Journal: bioRxiv

Article Title: AXL-GAS6/PROS1 Interaction: A Critical Switch Between Aberrant- and Healthy Repair Following Alveolar Lung Injury

doi: 10.1101/2025.04.07.647567

Figure Lengend Snippet: AXL is predominantly expressed in basal and aberrant basaloid cells, promoting proliferation via AXL-GAS6 signaling. (A) UMAP plots visualize AXL and its ligands, GAS6 and PROS1 normalized expression in the epithelial cell compartment of an integrated scRNA-seq IPF atlas. (B) Quantification of AXL and its ligands expression in epithelial cells of IPF patients and the non-diseased controls. (C) Expression of AXL and its ligands within the epithelial cells compartment in the lung of PF patients and the non-diseased controls. (D-E) AXL and its ligands expression in SAEC under submerged culture condition. (F) Representative western blot image validating knockout of AXL via CRISPR/Cas9 in 2 different donors. AXL (140 KDa, red bands), loading control ß-Actin (42 KDa, green bands), and M is protein marker. (G) Quantification of AXL expressions of each donor before and after CRISPR/Cas9 mediated knockdown. Experiments were done in duplicate. (H) Proliferation of AXL WT and AXL KD SAEC as assessed by BrdU in all donors 48 hours post seeding. (I) Correlation between AXL expression in (G) and SAEC proliferation in (H). (J) Proliferation of AXL WT and (K) AXL KD SAEC as assessed by BrdU 48 hours post treatment with rhGAS6/ rhPROS1/ combination of both proteins. WT: Wildtype; KD: Knockdown. Data are shown as mean ± SEM of n= 4 – 7. P > 0.05 (ns/ non-significant); P ≤ 0.05 (*); P ≤ 0.001 (***).

Article Snippet: GAS6 was measured in human BALF or SAEC supernatant using the human GAS6 DuoSet ELISA kit (R&D Systems, #DY885B) and the human PROS1 ELISA Kit (Aviva Systems Biology, #OKBB010207).

Techniques: Expressing, Western Blot, Knock-Out, CRISPR, Control, Marker, Knockdown

Journal: bioRxiv

Article Title: AXL-GAS6/PROS1 Interaction: A Critical Switch Between Aberrant- and Healthy Repair Following Alveolar Lung Injury

doi: 10.1101/2025.04.07.647567

Figure Lengend Snippet: (A) SAEC basal cells were allowed to differentiate for 23 days. On day 23 post ALI, 5 ng/ml rhTGF-ß was added to the culture. On day 26 post ALI, rhTGF-ß was added together with rhGAS6, rhPROS1, or a combination of both. Analysis was performed on day 29 post ALI. Scheme was created in BioRender. Geillinger-kästle, K. (2025) https://BioRender.com/k46g355 . (B) GAS6 and (C) PROS1 concentration in the SAEC cell culture supernatant post rhTGF-ß treatment compared to the BSA control. (D) Epithelial barrier integrity as measured by FITC-dextran permeability assays. (E) Fold change of AXL mRNA expression as measured by qPCR. (F) Correlation between slope measured by FITC- dextran permeability assay (D) and ΔCT of AXL (r= -0.9519 R 2 = 0.9062) measured by qPCR (E). Data are shown as mean ± SEM of n= 4 – 5. P > 0.05 (ns/ non-significant); P ≤ 0.05 (*); P ≤ 0.01 (**).

Article Snippet: GAS6 was measured in human BALF or SAEC supernatant using the human GAS6 DuoSet ELISA kit (R&D Systems, #DY885B) and the human PROS1 ELISA Kit (Aviva Systems Biology, #OKBB010207).

Techniques: Concentration Assay, Cell Culture, Control, Permeability, Expressing, FITC-Dextran Permeability Assay

Journal: bioRxiv

Article Title: AXL-GAS6/PROS1 Interaction: A Critical Switch Between Aberrant- and Healthy Repair Following Alveolar Lung Injury

doi: 10.1101/2025.04.07.647567

Figure Lengend Snippet: After injury, the normal repair process involves an initial inflammatory response, followed by AXL- GAS6 interaction, which promotes cell proliferation. Once sufficient proliferation is achieved, AXL-PROS1 binding occurs, halting proliferation and triggering cell differentiation. This balance between proliferation and differentiation ensures proper repair, leading to healthy alveoli. However, dysbalanced AXL signaling due to repeated injury and persistently high TGF- ß levels, can result in aberrant repair and irreversible fibrosis (IPF). This may occur due to high-affinity interaction between AXL and GAS6 prevents PROS1 from effectively binding to AXL. As a result, excessive proliferation occurs with insufficient differentiation, leading to aberrant repair and fibrosis. Scheme was created in BioRender. Geillinger-kästle, K. (2025) https://BioRender.com/q77c168.xss

Article Snippet: GAS6 was measured in human BALF or SAEC supernatant using the human GAS6 DuoSet ELISA kit (R&D Systems, #DY885B) and the human PROS1 ELISA Kit (Aviva Systems Biology, #OKBB010207).

Techniques: Binding Assay, Cell Differentiation

Journal: Cancer Science

Article Title: AXL‐Mediated Drug Resistance in ALK‐Rearranged NSCLC Enhanced by GAS6 From Macrophages and MMP11 Positive Fibroblasts

doi: 10.1111/cas.70006

Figure Lengend Snippet: AXL upregulation was observed in patient‐derived cell lines established after alectinib resistance. (A) Volcano plot of 58 receptor tyrosine kinase expressions in two patient‐derived cell lines (PDCs), JFCR‐028‐3 and JFCR‐028‐4, analyzed via RNA sequencing. (B) Histogram showing the AXL expression levels in the two PDCs analyzed via flow cytometry. The light blue and orange lines denote the isotype controls (mouse IgG, mIgG). (C) Immunoblot analysis of phosphor‐AXL (pAXL), and AXL in the two PDCs. (D) Bar graphs of enzyme‐linked immunosorbent assay (ELISA) showing the GAS6 concentrations (ng/mL) in the pleural effusion from which the two PDCs were established. Similar experiments were conducted twice (B, C) or thrice (D), and representative data are shown.

Article Snippet: Enzyme‐linked immunosorbent assay (ELISA) was conducted using Human Gas6 DuoSet ELISA (R&D Systems) and DuoSet Ancillary Reagent Kit 2 (R&D Systems) according to the manufacturer's protocol.

Techniques: Derivative Assay, RNA Sequencing, Expressing, Flow Cytometry, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Cancer Science

Article Title: AXL‐Mediated Drug Resistance in ALK‐Rearranged NSCLC Enhanced by GAS6 From Macrophages and MMP11 Positive Fibroblasts

doi: 10.1111/cas.70006

Figure Lengend Snippet: The overexpression of AXL and the presence of GAS6 were confirmed to have contributed to the development of resistance to alectinib, as evidenced by in vitro and in vivo studies. (A) Immunoblot analysis of phosphor‐AXL (pAXL) and AXL in H3122 parental and AXL‐overexpressing (AXL) cells. (B, E) Bar graphs showing the effects of alectinib (100 nmol/L) on the viability of H3122 parental (PT) and AXL‐overexpressing (AXL) cells. Recombinant GAS6 (400 ng/mL) (B) or conditioned medium from parental (PT) and GAS6‐overexpressing (GAS6) NIH3T3 single‐clone cells (no. 37) (E) was added 6 h before treatment. Cell viability was evaluated after 72 h using the CellTiter‐Glo Assay. (C) Immunoblot analysis of GAS6 in parental NIH3T3 cells as well as polyclonal (GAS6 poly) and single‐clone (GAS6 #37) GAS6‐overexpressing NIH3T3 cells. (D) Bar graphs of enzyme‐linked immunosorbent assay (ELISA) showing the concentrations (ng/mL) of GAS6 in the conditioned medium of parental (PT) NIH3T3 cells as well as polyclonal (GAS6 poly) and single‐clone (GAS6 no. 37) GAS6‐overexpressing NIH3T3 cells. An asterisk (*) indicates a value below the detection threshold. (F) Parental (PT) and AXL‐overexpressing (AXL) H3122 cells and parental (PT) and human GAS6‐overexpressing (GAS6) NIH3T3 polyclonal cells were subcutaneously transplanted into BALB/c nu/nu mice. The NIH3T3 cells were repeatedly transplanted every 3 days. Once the average tumor volume reached approximately 200 mm 3 , the mice were treated with vehicle or alectinib (30 mg/kg) once daily for 5 days per week via oral gavage ( n = 6). * p < 0.05, *** p < 0.001, **** p < 0.0001 (one‐way analysis of variance following Bonferroni's multiple comparisons test). Similar experiments were conducted twice (A–C, E) or thrice (D), and representative data are shown.

Article Snippet: Enzyme‐linked immunosorbent assay (ELISA) was conducted using Human Gas6 DuoSet ELISA (R&D Systems) and DuoSet Ancillary Reagent Kit 2 (R&D Systems) according to the manufacturer's protocol.

Techniques: Over Expression, In Vitro, In Vivo, Western Blot, Recombinant, Glo Assay, Enzyme-linked Immunosorbent Assay

Journal: Cancer Science

Article Title: AXL‐Mediated Drug Resistance in ALK‐Rearranged NSCLC Enhanced by GAS6 From Macrophages and MMP11 Positive Fibroblasts

doi: 10.1111/cas.70006

Figure Lengend Snippet: Gilteritinib was effective in overcoming resistance to ALK‐TKI treatment resulting from the activation of the GAS6/AXL signaling pathway. (A, B) Bar graphs showing the effects of gilteritinib (100 nmol/L) on the viability of H3122 parental (PT) and AXL‐overexpressing (AXL) cells. Recombinant GAS6 (400 ng/mL) (A) or conditioned medium from parental (PT) and single‐clone (#37) GAS6‐overexpressing (GAS6) NIH3T3 cells (B) was added 6 h before drug treatment. Cell viability was evaluated after 72 h using the CellTiter‐Glo Assay. (C) Immunoblot analysis of ALK, AXL, PARP, and the downstream pathways of ALK in parental (PT) and AXL‐overexpressing (AXL) H3122 cells. The cells were treated with the indicated concentrations of the drugs (Ale, alectinib; Gil, gilteritinib) for 6 h. Recombinant GAS6 (100 ng/mL) was added 1 h before the initiation of drug treatment.

Article Snippet: Enzyme‐linked immunosorbent assay (ELISA) was conducted using Human Gas6 DuoSet ELISA (R&D Systems) and DuoSet Ancillary Reagent Kit 2 (R&D Systems) according to the manufacturer's protocol.

Techniques: Activation Assay, Recombinant, Glo Assay, Western Blot

Journal: Cancer Science

Article Title: AXL‐Mediated Drug Resistance in ALK‐Rearranged NSCLC Enhanced by GAS6 From Macrophages and MMP11 Positive Fibroblasts

doi: 10.1111/cas.70006

Figure Lengend Snippet: Gas6 expression in the host nontumor cells was elevated from the early phase of alectinib treatment. (A) A schematic diagram of the methodology of the in vivo experiment. The tumor size at the specified time point is indicated by the line graphs. The timing of tumor collection is indicated by the arrows. BALB/c‐nu, BALB/c‐nude mice; H3122‐AXL, AXL‐overexpressing H3122 cells; RNA‐seq, RNA‐sequencing; RT‐qPCR, Quantitative reverse transcription‐polymerase chain reaction. (B, C) Hierarchical clustering with a heatmap of the bulk RNA sequencing (RNA‐seq) data, with the human (B) and murine (C) genomes used as the references. The timing of tumor collection (day) and the type of treatment (alectinib, ALE; vehicle, VEH) are indicated at the top. (D, E) Bar graphs of RT‐qPCR showing the expression levels of murine Gas6 (mGas6) and human GAS6 (hGAS6) in tumor samples obtained at various time points throughout the treatment period. hACTB, Human ACTB; mActb, Murine Actb. (F) Bar graphs of the transcript per million (TPM) value of mGas6 and hGAS6 determined via RNA‐seq analysis. Each data point represents a single TPM value derived from the bulk tumor sample collected on days 2–11. The number of samples at each time point is 3. * p < 0.05 (unpaired t ‐test)

Article Snippet: Enzyme‐linked immunosorbent assay (ELISA) was conducted using Human Gas6 DuoSet ELISA (R&D Systems) and DuoSet Ancillary Reagent Kit 2 (R&D Systems) according to the manufacturer's protocol.

Techniques: Expressing, In Vivo, RNA Sequencing, Quantitative RT-PCR, Reverse Transcription, Polymerase Chain Reaction, Derivative Assay

Journal: Cancer Science

Article Title: AXL‐Mediated Drug Resistance in ALK‐Rearranged NSCLC Enhanced by GAS6 From Macrophages and MMP11 Positive Fibroblasts

doi: 10.1111/cas.70006

Figure Lengend Snippet: Alectinib treatment resulted in an increase in the number of macrophages and resulting in elevated Gas6 levels in the tumor microenvironment (TME). (A) Bar plots showing the proportion of macrophages in samples collected from days 2 to 8 (left). Pie chart illustrating the proportion of immune cells in the day 8 samples. The mean proportions from triplicate samples are shown (right). B Cells Naïve, naïve B cells; DC Immature, immature dendritic cells; NK Activated, activated natural killer cells. (B) t‐SNE plots of 23,920 cells colored by sample origin (left) and 10 clusters determined using the k‐means method (right). Each cluster was annotated on the basis of differentially upregulated genes and expression levels of cell markers. “Veh_d8_1” represents replicate 1 of vehicle‐treated sample collected on day 8. Ale, alectinib; Veh, vehicle. (C) t‐SNE plots illustrating the expression patterns of Gas6 (left) and violin plots showing the Gas6 expression levels (log 2 UMI count) in each cluster (right). (D) t‐SNE plots of cluster 4 colored by treatment group (left), violin plots showing the Gas6 expression (log 2 UMI count) in each group (middle), and a pie chart illustrating the proportion of each group (right). * p < 0.05 (unpaired t ‐test)

Article Snippet: Enzyme‐linked immunosorbent assay (ELISA) was conducted using Human Gas6 DuoSet ELISA (R&D Systems) and DuoSet Ancillary Reagent Kit 2 (R&D Systems) according to the manufacturer's protocol.

Techniques: Expressing

Journal: Cancer Science

Article Title: AXL‐Mediated Drug Resistance in ALK‐Rearranged NSCLC Enhanced by GAS6 From Macrophages and MMP11 Positive Fibroblasts

doi: 10.1111/cas.70006

Figure Lengend Snippet: Alectinib treatment resulted in alterations in the expression of cancer‐associated fibroblasts, resulting in elevated Gas6 levels in the tumor microenvironment (TME). (A) t‐SNE plots of clusters 5 and 6 colored by treatment group (left), violin plots showing the Gas6 expression (log 2 UMI count) in each group (middle), and a pie chart illustrating the proportion of each group (right). (B) Violin plots showing the Mmp11 expression (log 2 UMI count) in each group. (C) t‐SNE plots of cluster 5 and 6 cells with high Gas6 expression (log 2 UMI count > 1), colored by Mmp11 and Gas6 expression levels (log 2 UMI count). (D) A schematic diagram of ALK‐tyrosine kinase inhibitor (ALK‐TKI) resistance by the increased GAS6 in the TME. CAFs, cancer‐associated fibroblasts.

Article Snippet: Enzyme‐linked immunosorbent assay (ELISA) was conducted using Human Gas6 DuoSet ELISA (R&D Systems) and DuoSet Ancillary Reagent Kit 2 (R&D Systems) according to the manufacturer's protocol.

Techniques: Expressing