Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Tylosema esculentum (Marama) Tuber and Bean Extracts Are Strong Antiviral Agents against Rotavirus Infection

doi: 10.1155/2011/284795

Figure Lengend Snippet: Dose-dependent cytopathic effect of rotavirus on bovine intestinal epithelial cells (CIEB), pig intestinal epithelial cells (CLAB), and human intestinal epithelial cells (H4). The graphs for cytopathic effect of rotavirus on the different cells are shown in terms of mean absorbance at each concentration ± standard deviation at 595 nm.

Article Snippet: NO 2 release was significant from human intestinal epithelial cells (H4) exposed to all extracts used {seed coat water and ethanolic extracts (MSCW and MSCE, resp.), cotyledon water and ethanolic extracts (MCW and MCE, resp.), and tuber water extract (MTW)}, while only MCE and MTW significantly increased NO in CLAB cells ( ).

Techniques: Concentration Assay, Standard Deviation

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Tylosema esculentum (Marama) Tuber and Bean Extracts Are Strong Antiviral Agents against Rotavirus Infection

doi: 10.1155/2011/284795

Figure Lengend Snippet: Survival of bovine intestinal epithelial cells (CIEB), pig intestinal epithelial cells (CLAB), and human intestinal epithelial cells (H4) from cytopathic effects of rotavirus in the presence of T. esculentum extracts. Shown above are bars of % survival of epithelial cells {% change in cell viability after coincubation of rotavirus with marama extracts compared to rotavirus only wells at OD 595 nm ± % standard deviation}, respective levels of significance in terms of T values (*). T values were determined using Statsoft Statistica software ( P < .05). Tests were done in triplicate wells. * T value between 0 and 5 (low level significance), ** T value greater than 5, but less than 10 (middle level significance), and *** T value greater than 10 (high level significance). MCE: T. esculentum bean cotyledon ethanolic extract, MSCE: T. esculentum bean husk ethanolic extract, MTW: T. esculentum tuber water extract, MSCW: T. esculentum bean husk water extract, MCW: T. esculentum bean cotyledon water extract, and Combivir: (lamivudine and zidovudine).

Article Snippet: NO 2 release was significant from human intestinal epithelial cells (H4) exposed to all extracts used {seed coat water and ethanolic extracts (MSCW and MSCE, resp.), cotyledon water and ethanolic extracts (MCW and MCE, resp.), and tuber water extract (MTW)}, while only MCE and MTW significantly increased NO in CLAB cells ( ).

Techniques: Standard Deviation, Software

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Tylosema esculentum (Marama) Tuber and Bean Extracts Are Strong Antiviral Agents against Rotavirus Infection

doi: 10.1155/2011/284795

Figure Lengend Snippet: T. esculentum extracts show direct inhibition of rotavirus infectivity on bovine intestinal epithelial cells (CIEB), pig intestinal epithelial cells (CLAB), and human intestinal epithelial cells (H4). Shown above are bars of % survival of epithelial cells {% change in cell viability after pre-exposure of rotavirus to T. esculentum extracts compared to rotavirus only wells at OD 595 nm ± % standard deviation}, respective levels of significance in terms of T values (*). T values were determined using Statsoft Statistica software ( P < .05). Tests were done in triplicate wells. * T value between 0 and 5 (low level significance), ** T value greater than 5, but less than 10 (middle level significance), and *** T value greater than 10 (high level significance). MCE: T. esculentum bean cotyledon ethanolic extract, MSCE: T. esculentum bean husk ethanolic extract, MTW: T. esculentum tuber water extract, MSCW: T. esculentum bean husk water extract, MCW: T. esculentum bean cotyledon water extract, and AZT: Combivir (lamivudine and zidovudine).

Article Snippet: NO 2 release was significant from human intestinal epithelial cells (H4) exposed to all extracts used {seed coat water and ethanolic extracts (MSCW and MSCE, resp.), cotyledon water and ethanolic extracts (MCW and MCE, resp.), and tuber water extract (MTW)}, while only MCE and MTW significantly increased NO in CLAB cells ( ).

Techniques: Inhibition, Infection, Standard Deviation, Software

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Tylosema esculentum (Marama) Tuber and Bean Extracts Are Strong Antiviral Agents against Rotavirus Infection

doi: 10.1155/2011/284795

Figure Lengend Snippet: Effect of coincubation of rotavirus with T. esculentum extracts on monolayer polarity (TER) (Ω/cm 2 ) of H4 cells over time. Shown graphs of change in epithelial cell monolayer polarity exposed to extracts or AZT and rotavirus (treatment wells), extracts or AZT alone (control), or rotavirus alone (negative control) over time compared to respective controls. Tests were done in triplicate wells (12-well Transwell plates). MCE: T. esculentum bean cotyledon ethanolic extract (1 mg/mL), MSCE: T. esculentum bean husk ethanolic extract (0.1 mg/mL), MTW: T. esculentum tuber water extract (0.1 mg/mL), MSCW: T. esculentum bean husk water extract (0.01 mg/mL), MCW: T. esculentum bean cotyledon water extract (2 mg/mL), and AZT: Combivir (lamivudine and zidovudine) (0.75 mg/mL).

Article Snippet: NO 2 release was significant from human intestinal epithelial cells (H4) exposed to all extracts used {seed coat water and ethanolic extracts (MSCW and MSCE, resp.), cotyledon water and ethanolic extracts (MCW and MCE, resp.), and tuber water extract (MTW)}, while only MCE and MTW significantly increased NO in CLAB cells ( ).

Techniques: Control, Negative Control

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Tylosema esculentum (Marama) Tuber and Bean Extracts Are Strong Antiviral Agents against Rotavirus Infection

doi: 10.1155/2011/284795

Figure Lengend Snippet: Effect of T. esculentum extracts on release of nitric oxide (NO) (% release compared to control) in human small intestine epithelial cells (H4) and pig small intestine epithelial cells (CLAB). MCE: T. esculentum bean cotyledon ethanolic extract (1 mg/mL), MSCE: T. esculentum bean husk ethanolic extract (0.1 mg/mL), MTW: T. esculentum tuber water extract (0.1 mg/mL), MSCW: T. esculentum bean husk water extract (0.01 mg/mL), and MCW: T. esculentum bean cotyledon water extract (2 mg/mL).

Article Snippet: NO 2 release was significant from human intestinal epithelial cells (H4) exposed to all extracts used {seed coat water and ethanolic extracts (MSCW and MSCE, resp.), cotyledon water and ethanolic extracts (MCW and MCE, resp.), and tuber water extract (MTW)}, while only MCE and MTW significantly increased NO in CLAB cells ( ).

Techniques: Control

Journal: Oncotarget

Article Title: Efficacy of a Mer and Flt3 tyrosine kinase small molecule inhibitor, UNC1666, in acute myeloid leukemia

doi:

Figure Lengend Snippet: (A) Chemical structure of UNC1666, with inhibition constant (K i ) of 0.16 nM for Mer (enzymatic IC 50 : 0.55 nM) and 0.67 nM for Flt3 (enzymatic IC 50 : 0.69 nM). (B) Chemical structure of UNC1653, which lacks significant activity against Mer (enzymatic IC 50 : 560 nM) and Flt3 (enzymatic IC 50 : 220 nM) and is used as a negative control in these studies. (C) Whole cell lysates from AML cell lines with known Flt3 mutation status were analyzed by immunoblot and demonstrate presence or absence of the Mer tyrosine kinase (above) and the Flt3 tyrosine kinase (middle). Actin is shown as an indicator of total protein (below).

Article Snippet: Additionally, UNC1666 inhibits Flt3 (MCE IC 50 0.69 nM; K i 0.67 nM) equipotently in enzymatic MCE assays.

Techniques: Inhibition, Activity Assay, Negative Control, Mutagenesis, Western Blot

Journal: Oncotarget

Article Title: Efficacy of a Mer and Flt3 tyrosine kinase small molecule inhibitor, UNC1666, in acute myeloid leukemia

doi:

Figure Lengend Snippet: (A) Mer was immunoprecipitated from AML cell lysates and phosphorylated ( p -Mer) and total Mer (~180 kDa) levels were assessed by immunoblot analysis. This representative blot of the Kasumi-1 cell line demonstrates decreased Mer phosphorylation after treatment with increasing doses of UNC1666. (B) Flt3 was immunoprecipitated from AML cell lysates and phosphorylated ( p -Flt3) and total Flt3 (130/160 kDa) levels were assessed by immunoblot analysis. This representative blot of the MV4;11 cell line demonstrates decreased Flt3 phosphorylation after treatment with increasing doses of UNC1666. (C) Inhibition of downstream signaling after administration of UNC1666 in a Mer expressing AML cell lines that does not express a Flt3-ITD mutation (Kasumi-1) compared with equivalent concentrations of vehicle (DMSO) or inactive control TKI UNC1653. Actin is shown as an indicator of total protein. (D) Downstream signaling after administration of UNC1666 in a Flt3-ITD AML cell line that does not express Mer (MV4;11). Representative blots from at least 3 independent experiments are shown. nM = nanomolar

Article Snippet: Additionally, UNC1666 inhibits Flt3 (MCE IC 50 0.69 nM; K i 0.67 nM) equipotently in enzymatic MCE assays.

Techniques: Immunoprecipitation, Western Blot, Phospho-proteomics, Inhibition, Expressing, Mutagenesis, Control

Journal: Oncotarget

Article Title: Efficacy of a Mer and Flt3 tyrosine kinase small molecule inhibitor, UNC1666, in acute myeloid leukemia

doi:

Figure Lengend Snippet: Mer pos or Flt3-ITD AML cell lines were treated with UNC1666, vehicle (DMSO), or inactive control TKI UNC1653 for 72 hours and then analyzed by flow cytometry after staining with YO-PRO-1 iodide and propidium iodide to identify apoptotic and dead cells. (A) Representative flow cytometry profiles of Kasumi-1 cells are shown. The percentages of live (lower left quadrant), early apoptotic (lower right quadrant), and late apoptotic/dead cells (upper quadrants) are shown. (B) Graphic representation of flow cytometric analyses of apoptotic/dead cells. Mean values and standard errors were derived from at least 3 independent experiments. * p < 0.05, *** p < 0.001, NS = not significant. (C) Cells were treated as indicated for 72 hours, whole cell lysates were prepared and the indicated apoptotic proteins were assessed by immunoblot analysis. Actin is shown as a loading control.

Article Snippet: Additionally, UNC1666 inhibits Flt3 (MCE IC 50 0.69 nM; K i 0.67 nM) equipotently in enzymatic MCE assays.

Techniques: Control, Flow Cytometry, Staining, Derivative Assay, Western Blot

Journal: Oncotarget

Article Title: Efficacy of a Mer and Flt3 tyrosine kinase small molecule inhibitor, UNC1666, in acute myeloid leukemia

doi:

Figure Lengend Snippet: Mer positive or Flt3-ITD AML cell lines were treated with UNC1666, vehicle (DMSO), or inactive control TKI UNC1653 for 72 hours, fixed with 100% ethanol and then analyzed by flow cytometry after staining with propidium iodide to identify stage of cell cycle. Graphic representation of cell cycle progress in Kasumi-1 and MV4;11 cells using ModFit analysis are shown. The percentages of cells in G2/M (light gray), S (white), and G1 (dark gray) phases are shown. Mean values and standard errors were derived from at least 3 independent experiments. * p < 0.05, ** p < 0.01.

Article Snippet: Additionally, UNC1666 inhibits Flt3 (MCE IC 50 0.69 nM; K i 0.67 nM) equipotently in enzymatic MCE assays.

Techniques: Control, Flow Cytometry, Staining, Derivative Assay

Journal: Oncotarget

Article Title: Efficacy of a Mer and Flt3 tyrosine kinase small molecule inhibitor, UNC1666, in acute myeloid leukemia

doi:

Figure Lengend Snippet: (A) Diagram of the replating assay. Cells were treated with UNC1666 or vehicle for 72 hours, then washed to remove any residual compound and equal numbers of viable cells (1.5 × 10 4 /ml) were replated in growth medium on Day 0. On Day 6 after replating, the number of viable cells was determined. (B) Graphic representation of the results of the replating assay in Mer or Flt3-ITD expressing cell lines demonstrating decreased rebound growth after treatment with UNC1666. Mean values and standard errors were derived from at least 3 independent experiments. (C) Colony-formation assays were performed using Mer or Flt3-ITD expressing AML cell lines. Cells were grown in soft agar with the indicated treatments. Graphic representation of reduced colony number after treatment with UNC1666, compared to vehicle or negative control TKI UNC1653. Mean values and standard errors were derived from at least 3 independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001, NS = not significant.

Article Snippet: Additionally, UNC1666 inhibits Flt3 (MCE IC 50 0.69 nM; K i 0.67 nM) equipotently in enzymatic MCE assays.

Techniques: Expressing, Derivative Assay, Negative Control

Journal: Oncotarget

Article Title: Efficacy of a Mer and Flt3 tyrosine kinase small molecule inhibitor, UNC1666, in acute myeloid leukemia

doi:

Figure Lengend Snippet: (A) Immunoblot analysis of Mer and Flt3 expression in lysates prepared from AML patient samples. (B) Flt3 mutation status of patient samples determined by molecular profiling. (C, D) Dose-dependent inhibition of Mer and Flt3 phosphorylation in response to treatment with UNC1666. AML blasts from patient sample #10510 (Mer positive, Flt3-ITD high allelic ratio) were treated with UNC1666 or vehicle for two hours. (C) Mer and Flt3 were immunoprecipitated from cell lysates and phosphorylated Mer ( p -Mer), total Mer (~180 kDa), phosphorylated Flt3 ( p -Flt3) and total Flt3 (130/160 kDa) were detected by immunoblot. (D) Phosphorylation of downstream signaling molecules was assessed by immunoblot after treatment with UNC1666 or vehicle.

Article Snippet: Additionally, UNC1666 inhibits Flt3 (MCE IC 50 0.69 nM; K i 0.67 nM) equipotently in enzymatic MCE assays.

Techniques: Western Blot, Expressing, Mutagenesis, Inhibition, Phospho-proteomics, Immunoprecipitation

Journal: Oncotarget

Article Title: Efficacy of a Mer and Flt3 tyrosine kinase small molecule inhibitor, UNC1666, in acute myeloid leukemia

doi:

Figure Lengend Snippet: (A) Graphic representation of apoptosis and cell death in AML patient samples after treatment with UNC1666 or vehicle for 72 hours. Apoptotic and dead cells were determined by flow cytometry after staining with YO-PRO-1 iodide and propidium iodide. Values derived from each sample are shown. (B) Colony-forming assays were performed in methylcellulose with the indicated treatments. Graphic representation of reduced colony number after treatment with UNC1666, compared to vehicle. Mean values and standard errors derived from triplicate samples are shown. (C) Graphic representation of the effect of UNC1666 on normal cord blood colony forming potential. Mean values and standard errors were derived from 3 independent experiments. ** p < 0.01, NS = not significant.

Article Snippet: Additionally, UNC1666 inhibits Flt3 (MCE IC 50 0.69 nM; K i 0.67 nM) equipotently in enzymatic MCE assays.

Techniques: Flow Cytometry, Staining, Derivative Assay

Journal: Inflammation

Article Title: FTO-dependent m 6 A Demethylation Activates Mxd1 To Enhance Vitamin D-induced Suppression of Neuroinflammation Via PTEN/AKT/PGC-1α Signaling Pathways in Microglia

doi: 10.1007/s10753-026-02450-5

Figure Lengend Snippet: VitD promotes M2 microglial polarization in LPS-stimulated BV-2 cells and mice. A-B CCK-8 assays of BV-2 cell viability following treatment with various concentrations of VitD, with or without LPS stimulation. Data are representative of three independent experiments and shown as mean ± SD. C Western blot analysis of CD16, CD18 (M1 markers), CD206 and ARG1 (M2 markers) in LPS- or LPS + VitD (VitD)-treated BV-2 cells and primary mouse microglia. Data are representative of three independent experiments and shown as mean ± SD. n = 3 ** P < 0.01. One-way ANOVA and Tukey’s test. D Immunofluorescence analysis of the levels of CD16 and CD206 in LPS- or LPS + VitD (VitD)-treated BV-2 cells. Scale bar: 50 μm. Right panel: Quantification of CD16 and CD206 IF intensity. Six regions were randomly selected for each group, and the IF intensities of CD16 and CD206 were quantified using the ImageJ software. Data are shown as mean ± SD. ** P < 0.01. One-way ANOVA and Tukey’s test. Red arrows indicate CD16-positive microglia and green arrows indicate CD206-positive microglia. E ELISA analysis of supernatant levels of TNF-α, IL1β, IL-6, IL-4 and IL-10 in LPS- or LPS + VitD (VitD)-treated BV-2 cells. n = 6. Data are shown as mean ± SD. ** P < 0.01. One-way ANOVA and Tukey’s test. F Immunofluorescence analysis of the levels of CD16 and CD206 in cerebral cortex and hippocampal CA1 area of LPS- or LPS + VitD(VitD)-treated mice. Scale bar: 20 μm. n = 6 mice per group. Data are shown as mean ± SD. ** P < 0.01. One-way ANOVA and Tukey’s test. Red arrows indicate CD16-positive microglia and green arrows indicate CD206-positive microglia.

Article Snippet: For immunofluorescence analysis, brain sections were cut at 7 μm, washed with 0.1 M PBS, permeabilized with 0.3% (v/v) Triton X-100-PBS, blocked with 10% (v/v) donkey serum for 1 h, and then incubated with primary antibodies against CD16 (1:200, ProteinTech, Wuhan, China), CD206 (1:200, RecordBio) at 4 °C overnight.

Techniques: CCK-8 Assay, Western Blot, Immunofluorescence, Software, Enzyme-linked Immunosorbent Assay

Journal: Inflammation

Article Title: FTO-dependent m 6 A Demethylation Activates Mxd1 To Enhance Vitamin D-induced Suppression of Neuroinflammation Via PTEN/AKT/PGC-1α Signaling Pathways in Microglia

doi: 10.1007/s10753-026-02450-5

Figure Lengend Snippet: Mxd1 is essential for VitD-induced microglial M2 polarization in LPS-stimulated conditions. A Immunohistochemistry analysis of Mxd1 expression in cerebral cortex and hippocampal CA1 area hippocampal CA1 area of mice receiving LPS or LPS + VitD (VitD) treatment. Scale bar: 20 μm. Quantification of Mxd1-positive area is shown in the right panel. n = 6 mice per group. Data are shown as mean ± SD. ** P < 0.01. One-way ANOVA and Tukey’s test. B Western blot analysis of the protein levels of Mxd1 and c-MYC in BV-2 cells treated with LPS or LPS + VitD (VitD). Data are representative of three independent experiments and shown as mean ± SD. n = 3. ** P < 0.01. One-way ANOVA and Tukey’s test. C Immunofluorescence analysis of the levels of Mxd1 in in BV-2 cells treated with LPS or LPS + VitD (VitD). Scale bar: 100 μm. Right panel: Quantification of Mxd1 IF intensity. Six regions were randomly selected for each group, and the IF intensity of Mxd1 was quantified using the ImageJ software. Data are shown as mean ± SD. ** P < 0.01. One-way ANOVA and Tukey’s test. D Western blot analysis of the protein levels of CD16, CD18, CD206, ARG1 and Mxd1 in LPS or LPS + VitD (VitD)-treated BV-2 cells with either negative control siRNA (siNC), or Mxd1 siRNA (siMxd1). Data are representative of three independent experiments and shown as mean ± SD. n = 3. ns: not significant; * P < 0.05, ** P < 0.01. Two-way ANOVA and Tukey’s test. E ELISA analysis of supernatant levels of TNF-α, IL1β, IL-6, IL-4 and IL-10 in LPS or VitD-treated BV-2 cells with siNC or siMxd1. Data are representative of three independent experiments and shown as mean ± SD. n = 3 ** P < 0.01. Two-way ANOVA and Tukey’s test.

Article Snippet: For immunofluorescence analysis, brain sections were cut at 7 μm, washed with 0.1 M PBS, permeabilized with 0.3% (v/v) Triton X-100-PBS, blocked with 10% (v/v) donkey serum for 1 h, and then incubated with primary antibodies against CD16 (1:200, ProteinTech, Wuhan, China), CD206 (1:200, RecordBio) at 4 °C overnight.

Techniques: Immunohistochemistry, Expressing, Western Blot, Immunofluorescence, Software, Negative Control, Enzyme-linked Immunosorbent Assay

Journal: Inflammation

Article Title: FTO-dependent m 6 A Demethylation Activates Mxd1 To Enhance Vitamin D-induced Suppression of Neuroinflammation Via PTEN/AKT/PGC-1α Signaling Pathways in Microglia

doi: 10.1007/s10753-026-02450-5

Figure Lengend Snippet: Effect of FTO on VitD-induced microglial M2 polarization. A Global m 6 A RNA methylation analysis of total mRNAs in LPS- and LPS + VitD(VitD)-treated BV-2 cells. Data are representative of three independent experiments and shown as mean ± SD. * P < 0.05; ** P < 0.01. One-way ANOVA and Tukey’s test. B Western blot of m 6 A methylation regulator (METTL3, METTL14, WTAP, ALKBH5, and FTO) in the same treatment conditions. Data are representative of three independent experiments and shown as mean ± SD. ns: not significant; ** P < 0.01. One-way ANOVA and Tukey’s test. C m 6 A quantification in BV-2 cells transfected with negative control siRNA (siNC) or FTO siRNA (siFTO) and treated with LPS or LPS + VitD (VitD). Data are representative of three independent experiments and shown as mean ± SD. * P < 0.05; ** P < 0.01. Two-way ANOVA and Tukey’s test. D Western blot of CD16, CD18, CD206, ARG1 and Mxd1 in the same treatment conditions. Data are representative of three independent experiments and shown as mean ± SD. ns: not significant; * P < 0.05; ** P < 0.01. Two-way ANOVA and Tukey’s test. E ELISA of cytokine levels in corresponding supernatants. Data are representative of three independent experiments and shown as mean ± SD. ns: not significant; ** P < 0.01. Two-way ANOVA and Tukey’s test. F Immunohistochemistry for Iba-1 in the cerebral cortex and hippocampal CA1 area of mice treated with vehicle, LPS, LPS + VitD, and LPS + VitD + FB23-2. Scale bar: 20 μm. Quantification data are shown in the right panel. n = 6 mice per group. Data are shown as mean ± SD. ** P < 0.01. One-way ANOVA and Tukey’s test.

Article Snippet: For immunofluorescence analysis, brain sections were cut at 7 μm, washed with 0.1 M PBS, permeabilized with 0.3% (v/v) Triton X-100-PBS, blocked with 10% (v/v) donkey serum for 1 h, and then incubated with primary antibodies against CD16 (1:200, ProteinTech, Wuhan, China), CD206 (1:200, RecordBio) at 4 °C overnight.

Techniques: Methylation, Western Blot, Transfection, Negative Control, Enzyme-linked Immunosorbent Assay, Immunohistochemistry

Journal: Inflammation

Article Title: FTO-dependent m 6 A Demethylation Activates Mxd1 To Enhance Vitamin D-induced Suppression of Neuroinflammation Via PTEN/AKT/PGC-1α Signaling Pathways in Microglia

doi: 10.1007/s10753-026-02450-5

Figure Lengend Snippet: PGC-1α is required for VitD-induced M2 microglial polarization. A Immunofluorescence analysis of PGC-1α nuclear localization in LPS- or LPS + VitD-treated BV-2 cells. Scale bar: 100 μm. Right panel: Quantification of PGC-1α IF intensity. Six regions were randomly selected for each group, and the IF intensity of PGC-1α was quantified using the ImageJ software. Data are shown as mean ± SD. ** P < 0.01. One-way ANOVA and Tukey’s test. B Western blot analysis of CD16, CD18, CD206 and ARG1 in LPS- or LPS + VitD(VitD)-treated BV-2 cells with vehicle or SR-18,292. Data are representative of three independent experiments and shown as mean ± SD. ns: not significant; ** P < 0.01. Two-way ANOVA and Tukey’s test. C ELISA quantification of TNF-α, IL1β, IL-6, IL-4 and IL-10 in culture supernatants from the same experimental groups. Data are representative of three independent experiments and shown as mean ± SD. ns: not significant; ** P < 0.01. Two-way ANOVA and Tukey’s test. D Immunohistochemical staining and quantification of Iba-1-positive cells in cerebral cortex and hippocampal CA1 area of LPS- or VitD-treated mice with vehicle or SR-18,292, and the quantification of Iba-1 positive cells in each group was shown in the low panel. Scale bar: 20 μm. n = 6 mice per group. Data are shown as mean ± SD. ** P < 0.01. One-way ANOVA and Tukey’s test.

Article Snippet: For immunofluorescence analysis, brain sections were cut at 7 μm, washed with 0.1 M PBS, permeabilized with 0.3% (v/v) Triton X-100-PBS, blocked with 10% (v/v) donkey serum for 1 h, and then incubated with primary antibodies against CD16 (1:200, ProteinTech, Wuhan, China), CD206 (1:200, RecordBio) at 4 °C overnight.

Techniques: Immunofluorescence, Software, Western Blot, Enzyme-linked Immunosorbent Assay, Immunohistochemical staining, Staining

Journal: Discover Oncology

Article Title: Single-cell profiling of ERBB family receptors identifies ERBB3 as a key regulator in head and neck squamous cell carcinoma progression

doi: 10.1007/s12672-025-03681-y

Figure Lengend Snippet: Cell–cell communication analysis reveals B cell-specific interaction rewiring in ERBB3-high tumors. Sample-wise composition of high vs. low ERBB3-expressing groups in an independent HNSCC scRNA-seq dataset.(B) Total number and average strength of inferred interactions across groups. (C) CellChat plots showing differential epithelial cell interactions; B cells exhibited the most dramatic rewiring. Red arrows indicate enhanced ligand–receptor interactions in the ERBB3-high group, whereas blue arrows indicate decreased interactions compared to the ERBB3-low group.(D–E) UMAP projection and further subclustering of B cells.(F) Bubble plot showing differential ligand–receptor interactions involving TNFRSF13B, IL4R, and IGHG between ERBB3-high and ERBB3-low groups. Each dot represents a ligand–receptor pair, where dot size indicates communication strength (interaction probability) and color intensity reflects statistical significance. Notably, signaling via TNFRSF13B, IL4R, and IGHG shows reduced interaction strength and significance in the ERBB3-high group. (G) Cell-type proportions in each group.(H) Pathway-specific interaction plots highlighting TNFRSF13B + B cell axes.(I) Heatmaps of differential interaction numbers and strengths.(J) Relative information flow of signaling pathways.(K) Dot plot of significant differential ligand–receptor pairs with p-values and communication probability

Article Snippet: Mice in the treatment group received intraperitoneal injections of anti-ERBB3 monoclonal antibody (MP-RM-1, MCE) at a dose of 10 mg/kg, twice per week for three consecutive weeks.

Techniques: Expressing, Protein-Protein interactions

Journal: Discover Oncology

Article Title: Single-cell profiling of ERBB family receptors identifies ERBB3 as a key regulator in head and neck squamous cell carcinoma progression

doi: 10.1007/s12672-025-03681-y

Figure Lengend Snippet: Prognostic modeling based on ERBB pathway activity and tumor microenvironment features. (A) Risk score distribution, survival status, and heatmap of ERBB family gene expression across TCGA patients.(B) Kaplan–Meier overall survival curves comparing high- and low-risk groups based on LASSO-derived ERBB prognostic signature.(C) Expression of ERBB3, EGFR, ERBB2, and ERBB4 between risk groups.(D) Time-dependent ROC curves showing prognostic performance of the risk model. The AUCs were 0.68 (1 year), 0.71 (3 years), and 0.74 (5 years).(E–H) Violin plots comparing ESTIMATE score, tumor purity, stromal score, and immune score between high- and low-risk groups.(I) Bar plot showing CIBERSORTx deconvolution of immune infiltration across groups.(J–K) Validation of risk model in GEO dataset, including survival curve and ROC analysis

Article Snippet: Mice in the treatment group received intraperitoneal injections of anti-ERBB3 monoclonal antibody (MP-RM-1, MCE) at a dose of 10 mg/kg, twice per week for three consecutive weeks.

Techniques: Activity Assay, Gene Expression, Derivative Assay, Expressing, Biomarker Discovery

Journal: Discover Oncology

Article Title: Single-cell profiling of ERBB family receptors identifies ERBB3 as a key regulator in head and neck squamous cell carcinoma progression

doi: 10.1007/s12672-025-03681-y

Figure Lengend Snippet: Prognostic value and clinical correlation of ERBB family members in HNSCC. (A–D) Kaplan–Meier curves for EGFR, ERBB2, ERBB3, and ERBB4; ERBB3-high patients show better overall survival ( p = 0.028).(E) Boxplot comparing ERBB3 expression in normal ( n = 44) and tumor tissues ( n = 520) from TCGA.(F) ERBB3 expression across pathological grades; higher expression observed in grade 4 tumors ( n = 7)

Article Snippet: Mice in the treatment group received intraperitoneal injections of anti-ERBB3 monoclonal antibody (MP-RM-1, MCE) at a dose of 10 mg/kg, twice per week for three consecutive weeks.

Techniques: Expressing

Journal: Discover Oncology

Article Title: Single-cell profiling of ERBB family receptors identifies ERBB3 as a key regulator in head and neck squamous cell carcinoma progression

doi: 10.1007/s12672-025-03681-y

Figure Lengend Snippet: ERBB3 knockdown impairs proliferation, colony formation, migration, and invasion in SCC9 cells. (A) ERBB3 protein levels across HNSCC cell lines based on DepMap portal.(B–C) Validation of ERBB3 knockdown by lentiviral shRNA via western blot and qRT-PCR.(D) Cell proliferation measured by CCK-8 assays over four days.(E–F) Representative images and quantification of colony formation assays.(G–H) Representative transwell migration assay and quantification.(I–J) Transwell invasion assay and quantification. Data are presented as mean ± SD; * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: Mice in the treatment group received intraperitoneal injections of anti-ERBB3 monoclonal antibody (MP-RM-1, MCE) at a dose of 10 mg/kg, twice per week for three consecutive weeks.

Techniques: Knockdown, Migration, Biomarker Discovery, shRNA, Western Blot, Quantitative RT-PCR, CCK-8 Assay, Transwell Migration Assay, Transwell Invasion Assay

Journal: Discover Oncology

Article Title: Single-cell profiling of ERBB family receptors identifies ERBB3 as a key regulator in head and neck squamous cell carcinoma progression

doi: 10.1007/s12672-025-03681-y

Figure Lengend Snippet: ERBB3 inhibition modulates immunosuppressive B cell phenotypes in vivo.、(A) Volcano plot showing differentially expressed genes between Scissor-high and Scissor-low tumor cell subsets.(B) KEGG enrichment showing suppression of antigen presentation pathways in ERBB3-high B cells.(C) Violin plot of CCDC50 expression in CD45⁺MS4A1⁺ B cells stratified by ERBB3 status.(D) UMAP projection of CCDC50 expression in single-cell data.(E) Flow cytometry showing reduced TNFRSF13B⁺CCDC50⁺ B cells upon ERBB3 inhibitor treatment.(F) Flow cytometry of MHC-II⁺ B cells showing enhanced antigen presentation following ERBB3 inhibition ( n = 3 mice per group). Data are shown as mean ± SEM. Statistical significance was determined using two-tailed unpaired Student’s t-test. * p < 0.05; ** p < 0.01, *** p < 0.001

Article Snippet: Mice in the treatment group received intraperitoneal injections of anti-ERBB3 monoclonal antibody (MP-RM-1, MCE) at a dose of 10 mg/kg, twice per week for three consecutive weeks.

Techniques: Inhibition, In Vivo, Immunopeptidomics, Expressing, Flow Cytometry, Two Tailed Test