Journal: Bioengineered

Article Title: Inhibin β-A (INHBA) induces epithelial–mesenchymal transition and accelerates the motility of breast cancer cells by activating the TGF-β signaling pathway

doi: 10.1080/21655979.2021.1957754

Figure Lengend Snippet: Primer sequences used for qRT-PCR in our study

Article Snippet: The plasmid containing human INHBA cDNA (OriGene, Beijing, China) or an empty vector as negative control (OriGene, Beijing, China) was transfected to MCF-7 cells.

Techniques:

Journal: Bioengineered

Article Title: Inhibin β-A (INHBA) induces epithelial–mesenchymal transition and accelerates the motility of breast cancer cells by activating the TGF-β signaling pathway

doi: 10.1080/21655979.2021.1957754

Figure Lengend Snippet: INHBA overexpression is associated with a poor survival rate in BC. (a) Upregulation of INHBA in BC tissue from TCGA database. (b) Upregulation of INHBA in two sub-types of BC as compared to normal tissues, using Oncomine database. (c) INHBA overexpression in BC patients with lymph node metastasis (N stage indicates the level of lymph node metastasis; the higher the number, the more metastasis in lymph node). Data were retrieved from TCGA database. (d) INHBA overexpression is correlated with a worse prognosis in BC patients (TCGA database). (e) BC data from Human Protein Atlas database was used analyze the correlation between the protein level of INHBA and the prognosis of BC patients ( P = 0.0041)

Article Snippet: The plasmid containing human INHBA cDNA (OriGene, Beijing, China) or an empty vector as negative control (OriGene, Beijing, China) was transfected to MCF-7 cells.

Techniques: Over Expression

Journal: Bioengineered

Article Title: Inhibin β-A (INHBA) induces epithelial–mesenchymal transition and accelerates the motility of breast cancer cells by activating the TGF-β signaling pathway

doi: 10.1080/21655979.2021.1957754

Figure Lengend Snippet: The INHBA is upregulated in BC cells and facilitates cell growth. (a) The INHBA overexpression in four BC cell lines was measured by RT-qPCR and Western blot. β-actin was used as the loading control. (b) The efficiency of INHBA knockdown in BT549 cells by sh-INHBA#1/2 or sh-NC vectors was assessed by RT-qPCR and Western blot. GAPDH was used as a loading control in Western blot. (c) CCK-8 assay and (d) colony formation assay were performed to examine the cell proliferation of BT549 cells after the transfection of shRNA. (e) The migration of transfected cells was examined by wound healing assay. (f) The invasion ability of transfected cells was determined by transwell assays (×200 magnification). Data are summary of 3 independent experiments (n = 3). The error bars are defined as s.d. *, P < 0.05, **, P < 0.01, and ***, P < 0.001

Article Snippet: The plasmid containing human INHBA cDNA (OriGene, Beijing, China) or an empty vector as negative control (OriGene, Beijing, China) was transfected to MCF-7 cells.

Techniques: Over Expression, Quantitative RT-PCR, Western Blot, Control, Knockdown, CCK-8 Assay, Colony Assay, Transfection, shRNA, Migration, Wound Healing Assay

Journal: Bioengineered

Article Title: Inhibin β-A (INHBA) induces epithelial–mesenchymal transition and accelerates the motility of breast cancer cells by activating the TGF-β signaling pathway

doi: 10.1080/21655979.2021.1957754

Figure Lengend Snippet: The overexpression of INHBA enhances BC cells migration and invasion. (a) The efficiency of INHBA overexpression in MCF-7 cells transfected by human INHBA cDNA or empty vectors were assessed by RT-qPCR and Western blot. GAPDH was used as a loading control in Western blot. (b) CCK-8 assay and (c) colony formation assay were performed to examine the cell proliferation of MCF-7 cells after the transfection of cDNA. (d) The migration of transfected cells was examined by wound healing assays. (e) The invasion of transfected cells was determined by transwell assays (×200 magnification). Data are summary of 3 independent experiments (n = 3). The error bars are defined as s.d. *, P < 0.05, **, P < 0.01, and ***, P < 0.001

Article Snippet: The plasmid containing human INHBA cDNA (OriGene, Beijing, China) or an empty vector as negative control (OriGene, Beijing, China) was transfected to MCF-7 cells.

Techniques: Over Expression, Migration, Transfection, Quantitative RT-PCR, Western Blot, Control, CCK-8 Assay, Colony Assay

Journal: Bioengineered

Article Title: Inhibin β-A (INHBA) induces epithelial–mesenchymal transition and accelerates the motility of breast cancer cells by activating the TGF-β signaling pathway

doi: 10.1080/21655979.2021.1957754

Figure Lengend Snippet: The overexpression of INHBA enhances the aggressiveness in MCF-10A cells. (a) The efficiency of INHBA overexpression in MCF-10A cells transfected by human INHBA cDNA or empty vectors were assessed by RT-qPCR and Western blot. GAPDH was used as a loading control in Western blot. (b) CCK-8 assay and (c) colony formation assay were performed to examine the cell proliferation of MCF-10A cells after INHBA overexpression. (d) The migration of transfected cells was examined by wound healing assays. (e) The invasion of transfected cells was determined by transwell assays (×200 magnification). Data are summary of 3 independent experiments (n = 3). The error bars are defined as s.d. *, P < 0.05, **, P < 0.01, and ***, P < 0.001

Article Snippet: The plasmid containing human INHBA cDNA (OriGene, Beijing, China) or an empty vector as negative control (OriGene, Beijing, China) was transfected to MCF-7 cells.

Techniques: Over Expression, Transfection, Quantitative RT-PCR, Western Blot, Control, CCK-8 Assay, Colony Assay, Migration

Journal: Bioengineered

Article Title: Inhibin β-A (INHBA) induces epithelial–mesenchymal transition and accelerates the motility of breast cancer cells by activating the TGF-β signaling pathway

doi: 10.1080/21655979.2021.1957754

Figure Lengend Snippet: INHBA induces the EMT in BC cells. (a) Western blot analysis of the EMT-related proteins after knockdown of INHBA in BT549 cells (left) and overexpression in MCF-7 cells (right). (b) RT-qPCR analysis of the EMT-related genes after knockdown of INHBA in BT549 cells (left) and overexpression in MCF-7 cells (right). GAPDH was used as a loading control in Western blot. Data are summary of 3 independent experiments (n = 3). The error bars are defined as s.d. *, P < 0.05, **, P < 0.01, and ***, P < 0.001

Article Snippet: The plasmid containing human INHBA cDNA (OriGene, Beijing, China) or an empty vector as negative control (OriGene, Beijing, China) was transfected to MCF-7 cells.

Techniques: Western Blot, Knockdown, Over Expression, Quantitative RT-PCR, Control

Journal: Bioengineered

Article Title: Inhibin β-A (INHBA) induces epithelial–mesenchymal transition and accelerates the motility of breast cancer cells by activating the TGF-β signaling pathway

doi: 10.1080/21655979.2021.1957754

Figure Lengend Snippet: INHBA expression is positively correlated with TGF-β1, Smad2, Smad7, Snail and Slug, and negatively correlated with CK-19 in BC tissues. Pearson correlation analysis between INHBA expression and several TGF-β and EMT related genes using a web-based predictive tool ENCORI ( http://starbase.sysu.edu.cn )

Article Snippet: The plasmid containing human INHBA cDNA (OriGene, Beijing, China) or an empty vector as negative control (OriGene, Beijing, China) was transfected to MCF-7 cells.

Techniques: Expressing

Journal: Bioengineered

Article Title: Inhibin β-A (INHBA) induces epithelial–mesenchymal transition and accelerates the motility of breast cancer cells by activating the TGF-β signaling pathway

doi: 10.1080/21655979.2021.1957754

Figure Lengend Snippet: INHBA overexpression activates the TGF-β signaling pathway, which is suppressed by TGF-β inhibitor. (a) The protein level of TGF-β1, Smad2, p-Smad2, Smad3, p-Smad3 and VEGF-A in BT549 (INHBA knockdown) and MCF-7 cells (INHBA overexpression). GAPDH was used as loading control in Western blot. (b) The protein level of Smad2, p-Smad2, Smad3, p-Smad3, TGF-β1, INHBA in MCT-7 cells treated by TGF-β inhibitor (SB-431,542). 10 µM SB-431,542 was used as final concentration in the medium. DMSO solvent was also used as a negative control. (c) The cell migration (upper two) and invasion (lower two) analyses for MCF-7 cells treated with (or without) TGF-β inhibitor (×200 magnification). (d) Western blot analysis of EMT-related genes in MCF-7 cells transfected with (or without) INHBA expression vector, along with (or without) SB-431,542. Data are summary of 3 independent experiments (n = 3). The error bars are defined as s.d. *, P < 0.05, **, P < 0.01, and ***, P < 0.001

Article Snippet: The plasmid containing human INHBA cDNA (OriGene, Beijing, China) or an empty vector as negative control (OriGene, Beijing, China) was transfected to MCF-7 cells.

Techniques: Over Expression, Knockdown, Control, Western Blot, Concentration Assay, Solvent, Negative Control, Migration, Transfection, Expressing, Plasmid Preparation

Journal: Bioengineered

Article Title: Inhibin β-A (INHBA) induces epithelial–mesenchymal transition and accelerates the motility of breast cancer cells by activating the TGF-β signaling pathway

doi: 10.1080/21655979.2021.1957754

Figure Lengend Snippet: In vivo tumorigenesis assay corroborates the oncogenic role of INHBA in BC cells. (a-b) MCF-7 cells transfected with empty vector, INHBA cDNA, sh-INHBA, or sh-NC was injected into RAG1-deficient mice (n = 6 mice in each group). The tumor volume and weight were measured for 7 weeks after tumor cell injection. (c) IHC staining of Ki-67 (cell proliferation marker) and INHBA in the tumor tissues of different experimental groups

Article Snippet: The plasmid containing human INHBA cDNA (OriGene, Beijing, China) or an empty vector as negative control (OriGene, Beijing, China) was transfected to MCF-7 cells.

Techniques: In Vivo, Transfection, Plasmid Preparation, Injection, Immunohistochemistry, Marker

Journal: Frontiers in Endocrinology

Article Title: Amelioration of polycystic ovarian morphology by Tokishakuyakusan in a PCOS rat model: association with bone morphogenetic protein 4

doi: 10.3389/fendo.2025.1649124

Figure Lengend Snippet: Bmp4 , Bmp5 , and Inhba expression in the ovaries. (A–C) Relative mRNA expression quantified by real-time PCR. Expression levels are relative to 18S ribosomal RNA. Data are shown as mean ± standard deviation with individual data points. P -values were obtained by one-way analysis of variance (ANOVA) with Tukey’s post hoc test (vehicle: n = 6; DHT: n = 9; DHT+TSS: n = 10). (D–G) Correlation analysis between gene expression and ovarian follicle counts in the DHT and DHT+TSS groups ( n = 6 per group). The correlation coefficient ( r 2 ) and P -value were calculated. DHT, 5α-dihydrotestosterone; TSS, Tokishakuyakusan; Bmp, bone morphogenetic protein; Inhba, inhibin-βa; PCR, polymerase chain reaction.

Article Snippet: Membranes were blocked for 1 h at room temperature with SuperBlock Blocking Buffer in TBS (37535; Thermo Fisher Scientific) and incubated overnight at 4°C with primary antibodies against BMP4 (MAB1049; 1:1,000, Merck Millipore, Burlington, MA, USA) or inhibin βA (17524-1-AP; 1:1,000, Proteintech Group, Inc., Rosemont, IL, USA).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Standard Deviation, Gene Expression, Polymerase Chain Reaction

Journal: Frontiers in Endocrinology

Article Title: Amelioration of polycystic ovarian morphology by Tokishakuyakusan in a PCOS rat model: association with bone morphogenetic protein 4

doi: 10.3389/fendo.2025.1649124

Figure Lengend Snippet: BMP4 and inhibin βA protein expression in the ovaries. (A) Representative Western blot images. (B, C) Quantification of BMP4 and inhibin βA protein levels relative to β-actin. Data are shown as mean ± standard deviation with individual data points (vehicle: n = 6; DHT: n = 9; DHT+TSS: n = 10). * P < 0.05, ** P < 0.01, one-way analysis of variance (ANOVA) followed by Tukey’s post hoc test; P < 0.0167 by Bonferroni correction. BMP, bone morphogenetic protein; DHT, 5α-dihydrotestosterone; TSS, Tokishakuyakusan.

Article Snippet: Membranes were blocked for 1 h at room temperature with SuperBlock Blocking Buffer in TBS (37535; Thermo Fisher Scientific) and incubated overnight at 4°C with primary antibodies against BMP4 (MAB1049; 1:1,000, Merck Millipore, Burlington, MA, USA) or inhibin βA (17524-1-AP; 1:1,000, Proteintech Group, Inc., Rosemont, IL, USA).

Techniques: Expressing, Western Blot, Standard Deviation

Journal: Frontiers in Endocrinology

Article Title: Amelioration of polycystic ovarian morphology by Tokishakuyakusan in a PCOS rat model: association with bone morphogenetic protein 4

doi: 10.3389/fendo.2025.1649124

Figure Lengend Snippet: Effect of Tokishakuyakusan (TSS) on target gene expressions in granulosa cells (GCs) derived from prenatally 5α-dihydrotestosterone (DHT)-treated rats. (A) Timeline of ovarian GC collection. (B, C) Relative mRNA expression of Bmp4 and Inhba in primary cultured GCs after 24 h of TSS treatment (125–500 μg/mL). Expression levels are relative to Gapdh . Data are shown as mean ± standard deviation with individual data points ( n = 3 per group). ** P < 0.01, P -values for the untreated control were obtained using one-way analysis of variance (ANOVA) followed by Dunnett’s post hoc test. PMSG, pregnant mare serum gonadotropin; Bmp, bone morphogenetic protein; Inhba, inhibin-βa; Gapdh, glyceraldehyde-3-phosphate dehydrogenase.

Article Snippet: Membranes were blocked for 1 h at room temperature with SuperBlock Blocking Buffer in TBS (37535; Thermo Fisher Scientific) and incubated overnight at 4°C with primary antibodies against BMP4 (MAB1049; 1:1,000, Merck Millipore, Burlington, MA, USA) or inhibin βA (17524-1-AP; 1:1,000, Proteintech Group, Inc., Rosemont, IL, USA).

Techniques: Derivative Assay, Expressing, Cell Culture, Standard Deviation, Control

Journal: eLife

Article Title: Fibrodysplasia ossificans progressiva mutant ACVR1 signals by multiple modalities in the developing zebrafish

doi: 10.7554/eLife.53761

Figure Lengend Snippet:

Article Snippet: Recombinant DNA reagent , Human ActivinA (plasmid) , Origene , Cat #RC203226 INHBA, inhibinβa, ActivinA , Cloned into pCS2 backbone.

Techniques: Sequencing, Recombinant, Plasmid Preparation, Clone Assay, Software

Journal: bioRxiv

Article Title: Integrin-TGFβ axis induces partial EMT in basal-like cells to lead collective invasion

doi: 10.1101/2025.04.04.647177

Figure Lengend Snippet: (A) Confocal imaging of ITGα2, Vimentin, and K8 in an IDC patient tissue section. Green contour: cluster of cancer cells with protrusive morphology; green arrowheads: basal-like cells (K8-low) at the tumor–stroma interface with high ITGα2 expression. White contour: cluster of cancer cells lacking basal-like cells (K8-high), with low ITGα2 expression; magenta arrowheads. White arrowheads: fibroblast-like cells (elongated, spindle-shaped). (B) Quantification of mean gray values for ITGα2 and Vimentin in basal-like (n = 33), luminal-like (n = 32), and fibroblast-like (n = 32) cells from one IDC patient tissue section. (C) Representative brightfield images of MMTV-PyMT organoids (ITGα2-WT or ITGα2-KO, gRNA1 and gRNA2) cultured in 3D Collagen I. Black arrowheads: invasive strands. (D) Percentage of organoids exhibiting one or more invasive strands in ITGα2-WT and ITGα2-KO (clones 1 and 2 from gRNA1) MMTV-PyMT organoids. (E) qPCR analysis of classical TGF-β and EMT target genes in ITGα2-WT and ITGα2-KO MMTV-PyMT organoids cultured in 3D Collagen I for three days. Values represent mean normalized mRNA expression (relative to housekeeping genes), shown for KO organoids relative to WT controls (dashed line). Data are presented as mean ± SD from three independent experiments. (F) Confocal imaging of Col ¾ and F-actin in ITGα2-WT and ITGα2-KO MMTV-PyMT organoids after one day in 3D Collagen I. (G, H) qPCR analysis of Vimentin and Slug mRNA expression in ITGα2-KO versus ITGα2-WT MMTV-PyMT organoids treated with Activin A (20 ng/μl) or vehicle control (0.1% BSA) for three days. Bar graphs represent mean normalized expression values ± SD from four independent experiments. (I) Kaplan–Meier analysis correlating high vs. low mRNA expression of INHBA, ITGA2, ITGB1, and their combinations (ITGA2 + ITGB1, or INHBA + ITGA2 + ITGB1) with distant metastasis-free survival (DMFS) in patients with grade 3 breast cancer. Scale bars: 100 μm (A, C), 50 μm (A, zoom-in), 50 μm (F), 10 μm (F, zoom-in). P values: two-sided unpaired Mann–Whitney test (E), two-sided Kruskal-Wallis test with Dunn’s multiple comparisons (G, H), Log-rank test (I).

Article Snippet: The following antibodies were used: rabbit anti-human Vimentin (Cat#ab92547, Abcam), chicken anti-human Vimentin (Cat#PA1-16759, Invitrogen) rabbit anti-human Keratin 14 (Cat# 905301, Biolegend), rat anti-mouse Keratin 8 (Cat# 531826, DSHB), rabbit anti-rat Collagen I cleavage site (Col ¾, Cat#0217-025, immunoGlobe), rabbit anti-human integrin α2 (Cat#ab181548, Abcam), rabbit anti-human Inhba (Cat#10651-1-AP, Proteintech), Mouse IgG1 isotype control (MAB002, R&D Systems), anti-Activin A antibody (Cat#AF338, R&D Systems).

Techniques: Imaging, Expressing, Cell Culture, Clone Assay, Control, MANN-WHITNEY

Journal: bioRxiv

Article Title: Integrin-TGFβ axis induces partial EMT in basal-like cells to lead collective invasion

doi: 10.1101/2025.04.04.647177

Figure Lengend Snippet: (A) DNA sequence of the Itgα2 gene to confirm gene knockout by Crispr-Cas9 gene editing. Red regions indicate the insertion of one base pair compared to the wildtype Itgα2 sequence. (B) Western blot analysis showing Itgα2 and GAPDH expression from whole cell lysates of MMTV-PyMT organoids WT or KO (gRNA1 or gRNA2). (C) Confocal imaging of Itgα2 and K8 in MMTV-PyMT organoids with Itgα2 WT or knockout (KO) grown in Collagen I for 3 days. White arrowheads: invading strands in Itgα2 WT organoids led by basal-like cells (low K8, high Itgα2, insets), White arrows: non-invading basal-like cells at the ECM interface in Itgα2 KO organoids (low K8, low Itgα2, insets). (D) Percentage of invasive organoids in MMTV-PyMT organoids Itgα2-WT versus Itgα2-KO (gRNA2). (E) Mean gray value of Col ¾ relative to Collagen I reflection in MMTV-PyMT organoids with Itgα2-WT and Itgα2-KO. Median: red lines, from n = 11 organoids per group from two independent experiments. (F) Single confocal slice showing INHBA and K8 expression in Itgα2-WT and Itgα2-KO MMTV-PyMT organoids cultured in 3D Collagen I for three days. Insets show basal-like cells (K8 low) guiding invasive strands (Itgα2-WT) or remaining at the organoid rim (Itgα2 KO, non-invasive). (G) Mean-gray value of Inhba in basal-like cells (K8-low) located at the rim of Itgα2 WT vs. KO MMTV-PyMT organoids (3D Collagen I, day 1). Median: red lines, n =11 cells from 6 Itgα2-WT organoids, and n = 12 cells from 7 Itgα2-KO organoids from one experiment. (H) qPCR analysis showing relative mRNA expression of CTGF in MMTV-PyMT Itgα2-KO organoids compared to Itgα2-WT organoids treated with Activin A ligand (20 ng/μl) or vehicle control (0.1% BSA) for three days. Bar graph represents mean normalized expression values (relative to housekeeping genes) ± SD from four independent experiments. (I) Kaplan-Meier plot correlating ITGB1 gene expression with DMFS in grade 3 breast cancer patients. Scale bars: 50 μm (C, F), 25 μm (C, F, Zoom in). P values, two-sided unpaired Mann–Whitney test (E, G), two-sided Kruskal-Wallis test (Dunn’s multiple comparison) (H), Logrank test (I).

Article Snippet: The following antibodies were used: rabbit anti-human Vimentin (Cat#ab92547, Abcam), chicken anti-human Vimentin (Cat#PA1-16759, Invitrogen) rabbit anti-human Keratin 14 (Cat# 905301, Biolegend), rat anti-mouse Keratin 8 (Cat# 531826, DSHB), rabbit anti-rat Collagen I cleavage site (Col ¾, Cat#0217-025, immunoGlobe), rabbit anti-human integrin α2 (Cat#ab181548, Abcam), rabbit anti-human Inhba (Cat#10651-1-AP, Proteintech), Mouse IgG1 isotype control (MAB002, R&D Systems), anti-Activin A antibody (Cat#AF338, R&D Systems).

Techniques: Sequencing, Gene Knockout, CRISPR, Western Blot, Expressing, Imaging, Knock-Out, Cell Culture, Control, Gene Expression, MANN-WHITNEY, Comparison