anti fbxo11  (Novus Biologicals)

Journal: Human Genetics and Genomics Advances

Article Title: Proteasomal activation ameliorates neuronal phenotypes linked to FBXO11 -deficiency

doi: 10.1016/j.xhgg.2025.100425

Figure Lengend Snippet: Generation of a neuronal FBXO11 deficient cell model using CRISPR-CAS9 (A) Outline of generation process of FBXO11 heterozygous and complete knockout hIPSC lines is shown, including validation of mutated clones and isogenic controls. (B) Representative western blot of all nine FBXO11 hIPSC lines is shown. Blots were stained with anti-FBXO11 and anti-GAPDH antibodies. (C) Quantification of three independent western blot experiments of FBXO11 hIPSC lines confirmed loss of FBXO11 in HET and KO lines. Individual values are shown as dots and mean values are shown as bars with SEM. Mean of all three WT controls was set to 1. p values were calculated using a one-sample t test with a hypothetical control mean set to 1. For all HET and KO lines compared with the control mean, reduction was significant at p < 0.01. (D) Schematic outline of differentiation protocol from hIPSCs to NPCs and neurons with timeline and culture media used. Below, experiments performed here are indicated at various timepoints. (E) Representative images of immunofluorescence of FBXO11 WT, HET, and KO NPCs stained with antibodies against neural progenitor markers Nestin (NES, red) and PAX6 (green) confirm differentiation to NPCs. Images of all nine NPC lines can be found in Figure S2 . Images were taken on an AxioImager Z2 with Apotome 3 with a 40× objective. Scale bar, 20 μm. (F) Quantification of PAX6-positive cells among NPCs shows comparable levels of PAX6-positive cells for WT, KO, and HET cells. Quantification for individual lines can be found in Figure S3 . For quantification, cells were analyzed using CellProfiler identifying DAPI-positive cells and PAX6 positive cells (fraction of PAX6-positive cells = PAX6 stained cells/DAPI-stained cells).

Article Snippet: Blots were stained with antibodies against FBXO11 (1:2,500, NB100-59826, Novus Bio), MAP2 (1:1,000), GAPDH (1:5,000, #2118, Cell Signaling), H3 (1:10,000, #4499, Cell Signaling), c-Myc (1:5,000, M4439, Sigma-Aldrich), c-Myc (1:2,500, #2272, Cell Signaling), FLAG (1:5,000, F7425, Sigma-Aldrich), FLAG (1:1,000, 194502, Addgene, gift from Melina Fan; http://n2t.net/addgene:194502 ; RRID: AB_2924869 ), and HA (1:2,000, H3663, Sigma-Aldrich).

Techniques: CRISPR, Knock-Out, Biomarker Discovery, Clone Assay, Western Blot, Staining, Control, Immunofluorescence

Journal: Human Genetics and Genomics Advances

Article Title: Proteasomal activation ameliorates neuronal phenotypes linked to FBXO11 -deficiency

doi: 10.1016/j.xhgg.2025.100425

Figure Lengend Snippet: Gene expression changes due to loss of FBXO11 in human neurons and fly heads (A) Principal-component analysis (PCA) of three FBXO11 WT and three KO neuron samples showed clear separation of WT and KO samples along the first principal component. (B) Enriched gene ontology (GO) terms among differentially expressed genes were grouped based on function and show a broad enrichment of biological processes involved in, e.g., development, signaling, and migration. Detailed results on individual enriched GO terms can be found in Figure S5 . (C) Integration of GO term analysis of FBXO11-deficient human neuron and Drosophila head transcriptome analysis. The top five biological processes enriched in GO term analysis of human neurons are shown in black. The enrichment of these processes in Fbxo11-deficient Drosophila heads are shown in green. (D) Stacked bar chart grouping genes expressed in FBXO11 KO neurons based on their differential gene expression and colored by corresponding expression changes during neuronal differentiation in a publicly available dataset on gene expression during differentiation from hIPSC to neurons. Increasing expression during differentiation is marked in green, and decreasing expression during differentiation is shown in pink. Unchanged expression is shown in gray. Number of genes with increasing expression during differentiation is increased for genes downregulated in FBXO11 KO neurons. down = downregulated, up = upregulated, not sig = expression not significantly changed, exp. = expression.

Article Snippet: Blots were stained with antibodies against FBXO11 (1:2,500, NB100-59826, Novus Bio), MAP2 (1:1,000), GAPDH (1:5,000, #2118, Cell Signaling), H3 (1:10,000, #4499, Cell Signaling), c-Myc (1:5,000, M4439, Sigma-Aldrich), c-Myc (1:2,500, #2272, Cell Signaling), FLAG (1:5,000, F7425, Sigma-Aldrich), FLAG (1:1,000, 194502, Addgene, gift from Melina Fan; http://n2t.net/addgene:194502 ; RRID: AB_2924869 ), and HA (1:2,000, H3663, Sigma-Aldrich).

Techniques: Gene Expression, Migration, Expressing

Journal: Human Genetics and Genomics Advances

Article Title: Proteasomal activation ameliorates neuronal phenotypes linked to FBXO11 -deficiency

doi: 10.1016/j.xhgg.2025.100425

Figure Lengend Snippet: Loss of FBXO11 alters neuronal migration, proliferation, and differentiation (A) Representative images of neurosphere assay on FBXO11 WT, HET, and KO NPCs imaged 48 h after plating. Inner circle represents initial neurosphere size, outer circle represents migration after 48 h. Images were taken on a Nikon Ts2-FL microscope. Scale bar, 100 μm. (B) Quantification of migration as ratio between area occupied at 48 h and plating (0 h). Migration is impacted in HET and more severely in KO neurospheres. Quantification of all nine individual lines can be found in Figure S7 A. At least 30 neurospheres per genotype (≥8 neurospheres per line) from three independent experiments were analyzed. Significance was calculated using a Student’s t test. (C) Representative images of immunofluorescence of FBXO11 WT, HET, and KO NPCs stained with antibodies against proliferation markers Ki-67 (red) and mitotic marker pHH3 (green) are shown. Images were taken on an AxioImager Z2 with a 20× objective. Scale bar, 100 μm. (D) Quantification of Ki67-positive cells among NPCs shows increased levels of Ki67-positive HET and KO cells. Quantification for individual lines can be found in Figure S7 D. For quantification, cells from 15 images were analyzed using CellProfiler identifying DAPI-positive and Ki67 positive cells (fraction of Ki67-positive cells = Ki67 stained cells/DAPI-stained cells). Significance was calculated using a Student’s t test. (E) Proliferation of differentiating neurons (D20-D28) was assessed using an XTT assay. Absorbance was normalized to the absorbance of D20 (NPC stage and first day of measurement). Plotted is the mean (circle) together with a trend line (colored and dashed) and the standard error (gray shading). Proliferation differences between WT vs. HET and HET vs. KO were significant for all three tested timepoints (D23, D26, D28, p < 0.01) and between WT vs. HET for two timepoints (D23 and D26, p < 0.01). The experiment was carried out three times with three technical replicates each. Significance was calculated using a Student’s t test. (F) Representative western blot of 3-week-old neurons (D42) stained against neuronal marker MAP2, FBXO11, and H3 as a loading control. (G) Quantification of MAP2 levels from western blot in (F) showed reduced MAP2 levels (normalized to loading control H3) for HET and KO neurons compared with WT. The experiment was performed three times. Mean expression of three WT controls were set to 1. Significance was calculated using a one-sample t test with a theoretical mean of 1. (H) Representative images of immunofluorescence of FBXO11 WT, HET, and KO 1-week old neurons (D28) stained with antibodies against neuronal markers MAP2 (red) and TUBB3 (green) are shown here and images of all nine neuronal lines can be found in Figure S4 . Images were taken on an AxioImager Z2 with a 40× objective. Scale bar, 40 μm. (I) Quantification of TUBB3-positive cells among neurons showed reduced levels of TUBB3-positive KO cells. Quantification for individual lines can be found in Figure S7 F. For quantification cells from at least 15 images were analyzed using CellProfiler identifying DAPI-positive cells (all cells) and TUBB3-positive cells (fraction of TUBB3-positive cells = TUBB3-stained cells/DAPI-stained cells). Significance was calculated using a Student’s t test. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: Blots were stained with antibodies against FBXO11 (1:2,500, NB100-59826, Novus Bio), MAP2 (1:1,000), GAPDH (1:5,000, #2118, Cell Signaling), H3 (1:10,000, #4499, Cell Signaling), c-Myc (1:5,000, M4439, Sigma-Aldrich), c-Myc (1:2,500, #2272, Cell Signaling), FLAG (1:5,000, F7425, Sigma-Aldrich), FLAG (1:1,000, 194502, Addgene, gift from Melina Fan; http://n2t.net/addgene:194502 ; RRID: AB_2924869 ), and HA (1:2,000, H3663, Sigma-Aldrich).

Techniques: Migration, Neurosphere Assay, Microscopy, Immunofluorescence, Staining, Marker, XTT Assay, Western Blot, Control, Expressing

Journal: Human Genetics and Genomics Advances

Article Title: Proteasomal activation ameliorates neuronal phenotypes linked to FBXO11 -deficiency

doi: 10.1016/j.xhgg.2025.100425

Figure Lengend Snippet: Deficiency of Fbxo11 leads to impaired behavior and dendritic branching in Drosophila melanogaster (A) Climbing assay upon pan-neuronal knockdown of Fbxo11 showed impaired locomotor ability for two of the three RNAi lines tested. Individual data points are shown as circles and summarized data are shown as boxplots. At least 200 flies in batches of 10 ( n = 20) were analyzed per condition. Significance was calculated using a Wilcoxon signed rank test. (B) Representative image of traced da neurons from control and knockdown larvae upon da neuron-specific knockdown (477-Gal4; UAS-mCD8GFP driver line) is shown. Images were acquired using a Zeiss LSM 710 confocal microscope with a 20× objective. Scale bar, 100 μm. (C) Quantification of total dendrite length of da neurons. (D) Quantification of number of branches from da neurons. Tracings of da neurons were performed in ImageJ using the NeuronJ plugin. At least 10 da neurons from five different larvae from two independent crosses were analyzed for each line. Statistical significance was calculated using a Student’s t test.

Article Snippet: Blots were stained with antibodies against FBXO11 (1:2,500, NB100-59826, Novus Bio), MAP2 (1:1,000), GAPDH (1:5,000, #2118, Cell Signaling), H3 (1:10,000, #4499, Cell Signaling), c-Myc (1:5,000, M4439, Sigma-Aldrich), c-Myc (1:2,500, #2272, Cell Signaling), FLAG (1:5,000, F7425, Sigma-Aldrich), FLAG (1:1,000, 194502, Addgene, gift from Melina Fan; http://n2t.net/addgene:194502 ; RRID: AB_2924869 ), and HA (1:2,000, H3663, Sigma-Aldrich).

Techniques: Climbing Assay, Knockdown, Control, Microscopy

Journal: Human Genetics and Genomics Advances

Article Title: Proteasomal activation ameliorates neuronal phenotypes linked to FBXO11 -deficiency

doi: 10.1016/j.xhgg.2025.100425

Figure Lengend Snippet: Rescue of FBXO11-deficiency-associated phenotypes with proteasome-activating substances (A) Formulas of tested substances PD169316, R-Verapamil, and Verapamil are shown. (B) Scoring scheme for rescue experiments corresponding to the level of completeness of the rescue. For dark-filled boxes, rescue resulted in almost complete normalization to control levels under the DMSO (75%–100%). For boxes filled with light shades of respective color, rescue levels reached 50%–75%. Different tested substances were supplemented to the fly food or the cell culture medium and are color-coded as follows: black – DMSO control, green – PD169316, red – R-Verapamil, purple – Verapamil. For fly experiments, all substances were used at 1 μM, for cell-based experiments, different concentrations were used (PD169316: 20 μM, R-Verapamil: 15 μM, Verapamil: 10 μM). (C) Climbing assay deficit upon pan-neuronal Fbxo11 knockdown with RNAi 1 could partially be rescued with proteasome-activating substances supplemented to the fly food. Improvement of phenotypes was seen when adding substances at time of egg laying (developmental supp.) or after flies hatched (adult supp.). At least 200 flies in batches of 10 ( n = 20) were analyzed per condition. (D) Total dendrite length of da neurons increased upon addition of proteasome-activating substances to the fly food at time of egg laying. At least five different neurons from five different larvae were analyzed per condition. (E and F) NPCs (E) or D28 neurons (F) were treated with proteasome-activating substances for 2 days (NPCs) or 1 week (neurons) before staining with Ki-67 antibody to assess proliferation. For quantification, cells from at least 15 images were analyzed using CellProfiler identifying DAPI-positive and Ki67-positive cells (% of Ki67-positive cells = Ki67-stained cells/DAPI-stained cells). For all plots, individual data points are shown as circles and summarized data are shown as boxplots. Statistical significance was calculated using either a Wilcoxon signed rank test (climbing assay) or a Student’s t test (da neuron assays and cell-based experiments) with correction for multiple testing. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: Blots were stained with antibodies against FBXO11 (1:2,500, NB100-59826, Novus Bio), MAP2 (1:1,000), GAPDH (1:5,000, #2118, Cell Signaling), H3 (1:10,000, #4499, Cell Signaling), c-Myc (1:5,000, M4439, Sigma-Aldrich), c-Myc (1:2,500, #2272, Cell Signaling), FLAG (1:5,000, F7425, Sigma-Aldrich), FLAG (1:1,000, 194502, Addgene, gift from Melina Fan; http://n2t.net/addgene:194502 ; RRID: AB_2924869 ), and HA (1:2,000, H3663, Sigma-Aldrich).

Techniques: Control, Cell Culture, Climbing Assay, Knockdown, Staining

Journal: Cancers

Article Title: FBXO11 Mediates Ubiquitination of ZEB1 and Modulates Epithelial-to-Mesenchymal Transition in Lung Cancer Cells

doi: 10.3390/cancers16193269

Figure Lengend Snippet: EMT is mainly induced in lung cancer cells by transcription factor ZEB1. ( A ) ZEB1 expression is increased in NiCl 2 -induced EMT. A549 and H1299 cells were treated with different concentrations of NiCl 2 for 2 d. Immunoblotting assays were performed to detect the expression of each EMT marker. ( B ) Knockdown of ZEB1 in cells resistant to the induction effect of NiCl 2 . A549 and H1299 shNC or shZEB1 cells were treated with 2 mM NiCl 2 , and changes in cell morphology were observed under a microscope. Scale bar: 100 μm. ( C ) FBXO11 is positively correlated with ZEB1. Correlation analysis of LUAD data for TCGA and GTEx on the GEPIA website showed a correlation coefficient of R = 0.69 ( p = 0; p < 0.01). TPM: transcripts per million. ( D ) FBXO11 exhibited higher expression in the normal tissue samples. The GEPIA database was used for FBXO11 expression in normal tissues and tumors (* p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: Membranes (Millipore, Shanghai, China, IPVH00010) were probed with primary antibodies, including FBXO11 (Proteintech, Wuhan, China, #67365-1-Ig), ZEB1 (Santa Cruz Biotechnology, Oregon, USA, #515797), E-cadherin (Proteintech, #20874-1-AP), N-cadherin (BD Transduction Laboratories, #610920), GFP (Proteintech, #66002-1-Ig), and GAPDH (GAPDH; Bioss, Woburn, MA, USA, #0978M).

Techniques: Expressing, Western Blot, Marker, Knockdown, Microscopy

Journal: Cancers

Article Title: FBXO11 Mediates Ubiquitination of ZEB1 and Modulates Epithelial-to-Mesenchymal Transition in Lung Cancer Cells

doi: 10.3390/cancers16193269

Figure Lengend Snippet: Correlation of FBXO11 expression with clinicopathological covariates in lung cancer patients.

Article Snippet: Membranes (Millipore, Shanghai, China, IPVH00010) were probed with primary antibodies, including FBXO11 (Proteintech, Wuhan, China, #67365-1-Ig), ZEB1 (Santa Cruz Biotechnology, Oregon, USA, #515797), E-cadherin (Proteintech, #20874-1-AP), N-cadherin (BD Transduction Laboratories, #610920), GFP (Proteintech, #66002-1-Ig), and GAPDH (GAPDH; Bioss, Woburn, MA, USA, #0978M).

Techniques: Expressing

Journal: Cancers

Article Title: FBXO11 Mediates Ubiquitination of ZEB1 and Modulates Epithelial-to-Mesenchymal Transition in Lung Cancer Cells

doi: 10.3390/cancers16193269

Figure Lengend Snippet: FBXO11 associates with ZEB1. ( A ) FBXO11 interacted with ZEB1 in a co-immunoprecipitation (co-IP) assay. HEK293T cells were transfected with Myc-tagged FBXO11 and Flag-tagged ZEB1 as indicated. Cell lysates were immunoprecipitated with either anti-Myc or anti-Flag antibodies and immunoblotted with anti-ZEB1 and anti-FBXO11 antibodies. ( B ) FBXO11 and ZEB1 co-location in the nucleus. Immunofluorescence assay probe colocalization of FBXO11 (green) and ZEB1 (red). Scale bar: 50 µm. ( C ) His pulldown assays showing FBXO11 bound to ZEB1 protein. A Coomassie blue staining image of PAGE gel is shown below to confirm the expression of pET28a and FBXO11. ( D ) His pulldown assays showing the CZF domain of ZEB1 bound to FBXO11 protein. A Coomassie blue staining image of PAGE gel is shown below to confirm the expression of various forms of ZEB1 proteins. ( E ) Molecular docking of FBXO11 protein and the ZEB1 CZF domain (725-1125 amino acids) truncated protein.

Article Snippet: Membranes (Millipore, Shanghai, China, IPVH00010) were probed with primary antibodies, including FBXO11 (Proteintech, Wuhan, China, #67365-1-Ig), ZEB1 (Santa Cruz Biotechnology, Oregon, USA, #515797), E-cadherin (Proteintech, #20874-1-AP), N-cadherin (BD Transduction Laboratories, #610920), GFP (Proteintech, #66002-1-Ig), and GAPDH (GAPDH; Bioss, Woburn, MA, USA, #0978M).

Techniques: Co-Immunoprecipitation Assay, Transfection, Immunoprecipitation, Immunofluorescence, Staining, Expressing

Journal: Cancers

Article Title: FBXO11 Mediates Ubiquitination of ZEB1 and Modulates Epithelial-to-Mesenchymal Transition in Lung Cancer Cells

doi: 10.3390/cancers16193269

Figure Lengend Snippet: FBXO11 stabilizes ZEB1 though ubiquitination activity. ( A ) FBXO11 promotes ZEB1 proteasomal degradation. ZEB1-Flag was cotransfected with FBXO11-Myc in HEK293T cells, together with GFP, as well as an empty vector, and treated with DMSO, MG132, or chloroquine as indicated. The expressions of ZEB1 and GFP were assessed by Western blot. ( B ) FBXO11 promotes K48 polyubiquitinated chain generation of ZEB1 protein. In cellular ubiquitination assays, FBXO11-Myc was co-transfected with ZEB1-Flag plasmids or with HA-Ub-K63 and HA-Ub-K48 plasmids. Western blot was performed, and cell lysates were immunoprecipitated with anti-Flag antibody, then detected by anti-Ub antibody to poly-Ub levels. ( C ) FBXO11 degrades ZEB1 protein in a concentration-dependent manner. HEK293T cells were transfected with ZEB1-Flag, GFP, or FBXO11-Myc for 48 h, and cell lysates were immunoblotted and screened with anti-Flag antibody. Quantities were calculated as fold changes of 0 μg FBXO11. Error bars represent the SD from 3 independent experiments. * indicates p < 0.05, and *** indicates p < 0.001. Each experiment was repeated at least three times. ( D ) Exogenous FBXO11 accelerates ZEB1 protein turnover. HEK293T cells were transfected with ZEB1-Flag and GFP, in combination with FBXO11-Myc, and treated with cycloheximide (CHX) as indicated. Cell lysates were subjected to western blot analysis with anti-ZEB1 and anti-GFP antibodies. The graph shows the quantification of ZEB1 protein levels (based on the band intensity from the gels) normalized to GFP over the time course. The ZEB1 protein level at the 0 h time point of CHX treatment was set as 100%. The experiment was repeated three times, and a representative experiment is presented.

Article Snippet: Membranes (Millipore, Shanghai, China, IPVH00010) were probed with primary antibodies, including FBXO11 (Proteintech, Wuhan, China, #67365-1-Ig), ZEB1 (Santa Cruz Biotechnology, Oregon, USA, #515797), E-cadherin (Proteintech, #20874-1-AP), N-cadherin (BD Transduction Laboratories, #610920), GFP (Proteintech, #66002-1-Ig), and GAPDH (GAPDH; Bioss, Woburn, MA, USA, #0978M).

Techniques: Ubiquitin Proteomics, Activity Assay, Plasmid Preparation, Western Blot, Transfection, Immunoprecipitation, Concentration Assay

Journal: Cancers

Article Title: FBXO11 Mediates Ubiquitination of ZEB1 and Modulates Epithelial-to-Mesenchymal Transition in Lung Cancer Cells

doi: 10.3390/cancers16193269

Figure Lengend Snippet: FBXO11 affects the expression of EMT-related factors. ( A ) Endogenous FBXO11 knockdown changes the expression of ZEB1 and EMT marker genes in lung adenocarcinoma cells. Immunoblotting analysis and quantitative RT-PCR analysis of ZEB1 and EMT markers in A549 and H1299 cells was transduced with lentiviral shRNAs targeting control or FBXO11. Error bars represent the SD from 3 independent experiments. * indicates p < 0.05, ** indicates p < 0.01, and *** indicates p < 0.001. ( B ) Immunofluorescence analysis of ZEB1 and E-cadherin protein expression in control and shFBXO11 of A549 and H1299 cells (ZEB1, red; E-cadherin, green; DAPI, blue). Scale bar: 50 μm. ( C ) FBXO11 overexpression affects changes cells into the mesenchymal phenotype. Cell morphology in A549 and H1299 cells with overexpressed Mock or FBXO11. Scale bars: 100 μm. ( D ) The overexpression of FBXO11 alters the expression of ZEB1 and EMT marker genes in lung adenocarcinoma cells. Immunoblotting and quantitative RT-PCR analysis of ZEB1 and EMT markers in A549 and H1299 cells transduced with Mock or FBXO11. Error bars represent the SD from 3 independent experiments. ** indicates p < 0.01, and *** indicates p < 0.001. ( E ) Wound-healing assay showing the migration of A549 and H1299 cells after transfection with Mock or FBXO11-Myc. Representative images are shown 0 and 48 h post scratch ( n = 3). Scale bars: 100 μm. ( F ) Transwell assay showing the invasion of A549 and H1299 cells transfected with Mock or FBXO11-Myc ( n = 3). Scale bars: 100 μm.

Article Snippet: Membranes (Millipore, Shanghai, China, IPVH00010) were probed with primary antibodies, including FBXO11 (Proteintech, Wuhan, China, #67365-1-Ig), ZEB1 (Santa Cruz Biotechnology, Oregon, USA, #515797), E-cadherin (Proteintech, #20874-1-AP), N-cadherin (BD Transduction Laboratories, #610920), GFP (Proteintech, #66002-1-Ig), and GAPDH (GAPDH; Bioss, Woburn, MA, USA, #0978M).

Techniques: Expressing, Knockdown, Marker, Western Blot, Quantitative RT-PCR, Transduction, Control, Immunofluorescence, Over Expression, Wound Healing Assay, Migration, Transfection, Transwell Assay

Journal: Cancers

Article Title: FBXO11 Mediates Ubiquitination of ZEB1 and Modulates Epithelial-to-Mesenchymal Transition in Lung Cancer Cells

doi: 10.3390/cancers16193269

Figure Lengend Snippet: The FBXO11–ZEB1 axis regulates the EMT pathway in LUAD. ( A ) Detection of FBXO11 or ZEB1 in A549 and H1299 cells expressing the indicated shRNAs. ( B ) The FBXO11–ZEB1 axis regulates epithelial–mesenchymal cell morphology in lung cancer cells. Morphological changes in A549 and H1299 control, shFBXO11, and shZEB1 and co-knockdown of ZEB1 and FBXO11 cells. Scale bars: 100 μm. ( C ) FBXO11 affects the migratory ability of A549 and H1299 cells via ZEB1. Scratching experiments were performed to analyze changes in the migratory capacity of A549 and H1299 control, shFBXO11, and shZEB1 and co-knockdown of ZEB1 and FBXO11 cells. Scale bar: 100 μm. The area of cell invasion was counted ( n = 6). ** indicates p < 0.01, *** indicates p < 0.001. ( D ) Cell invasiveness is moderated in vitro via the FBXO11–ZEB1 axis. A transwell assay to was conducted to analyze changes in the invasive capacity of A549 and H1299 control, shFBXO11, and shZEB1 and co-knockdown of ZEB1 and FBXO11 cells. Scale bar: 100 μm. The count of cells crossing the basement membrane was n = 6. ** indicates p < 0.01, and *** indicates p < 0.001.

Article Snippet: Membranes (Millipore, Shanghai, China, IPVH00010) were probed with primary antibodies, including FBXO11 (Proteintech, Wuhan, China, #67365-1-Ig), ZEB1 (Santa Cruz Biotechnology, Oregon, USA, #515797), E-cadherin (Proteintech, #20874-1-AP), N-cadherin (BD Transduction Laboratories, #610920), GFP (Proteintech, #66002-1-Ig), and GAPDH (GAPDH; Bioss, Woburn, MA, USA, #0978M).

Techniques: Expressing, Control, Knockdown, In Vitro, Transwell Assay, Membrane

Journal: Cancers

Article Title: FBXO11 Mediates Ubiquitination of ZEB1 and Modulates Epithelial-to-Mesenchymal Transition in Lung Cancer Cells

doi: 10.3390/cancers16193269

Figure Lengend Snippet: FBXO11 inhibits LUAD progression by stabilizing ZEB1. ( A ) Diagram of mouse hindlimb injection with A549 cells. ( B ) The FBXO11–ZEB1 axis controls tumor cell infiltration. Xenograft experiments were performed to analyze changes in the invasive ability of control, shFBXO11, shZEB1, co-knockdown, ZEB1, and FBXO11 cells in mice. Scale bars: 50 μm. ( C ) Xenograft experiments were performed to analyze changes in the invasive ability of A549, Mock, and FBXO11 cells in mice. Scale bars: 50 μm. ( D ) High ZEB1 and low FBXO11 expression predict poor prognosis. Kaplan–Meier plot of overall survival of lung cancer patients stratified by the expression of ZEB1 and FBXO11 genes. Data were obtained from KMplot.com.

Article Snippet: Membranes (Millipore, Shanghai, China, IPVH00010) were probed with primary antibodies, including FBXO11 (Proteintech, Wuhan, China, #67365-1-Ig), ZEB1 (Santa Cruz Biotechnology, Oregon, USA, #515797), E-cadherin (Proteintech, #20874-1-AP), N-cadherin (BD Transduction Laboratories, #610920), GFP (Proteintech, #66002-1-Ig), and GAPDH (GAPDH; Bioss, Woburn, MA, USA, #0978M).

Techniques: Injection, Control, Knockdown, Expressing

Journal: Cancers

Article Title: FBXO11 Mediates Ubiquitination of ZEB1 and Modulates Epithelial-to-Mesenchymal Transition in Lung Cancer Cells

doi: 10.3390/cancers16193269

Figure Lengend Snippet: FBXO11 regulates EMT and metastasis through ZEB1 in LUAD. FBXO11-dependent ubiquitination and ZEB1 protein stability regulate the invasion and metastasis of lung cancer cells.

Article Snippet: Membranes (Millipore, Shanghai, China, IPVH00010) were probed with primary antibodies, including FBXO11 (Proteintech, Wuhan, China, #67365-1-Ig), ZEB1 (Santa Cruz Biotechnology, Oregon, USA, #515797), E-cadherin (Proteintech, #20874-1-AP), N-cadherin (BD Transduction Laboratories, #610920), GFP (Proteintech, #66002-1-Ig), and GAPDH (GAPDH; Bioss, Woburn, MA, USA, #0978M).

Techniques: Ubiquitin Proteomics