Journal: SAGE Open Medicine

Article Title: Exploring the prognostic molecular mechanisms of medulloblastoma through methylation–transcriptome integration

doi: 10.1177/20503121251386139

Figure Lengend Snippet: Expression of INHBB and USP2 in medulloblastoma. (a) mRNA expression of INHBB and USP2 in three human cell lines. (b) IHC scoring results of INHBB and USP2 in medulloblastoma patients. (c) Expression of INHBB and USP2 in different subtypes of medulloblastoma. (d) Expression of INHBB and USP2 across four molecular subgroups. Box plots show significantly higher expression in Group 3 and Group 4 compared to WNT and SHH subtypes (Wilcoxon test, p < 2.22e-16). INHBB: inhibin beta B; USP2: ubiquitin-specific peptidase 2.

Article Snippet: Specific antibodies were utilized for inhibin beta B (INHBB; 1:400; Bioss, Bioss Antibodies, Woburn, MA, USA) and ubiquitin-specific peptidase 2 (USP2; 1:200; Proteintech (Proteintech Group, Rosemont, IL, USA)).

Techniques: Expressing, Ubiquitin Proteomics

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: Oncology Research

Article Title: INHBA Promotes the Progression of Gastric Cancer by Activating MAPK Signaling Pathway via Targeting ITGA6

doi: 10.32604/or.2025.070333

Figure Lengend Snippet: ITGA6 is the regulatory target of INHBA. ( A ) Differential expression genes (DEGs) heat map identified by RNA-seq. ( B ) The number of DEGs identified by RNA-seq. ( C ) Overlap analysis of RNA-seq and STAD-DEGs-up identified genes. ( D ) Expression of ITGA6 in GC paired tissue cohort from the GEPIA2 database. ( E ) Subcellular localization of ITGA6 from GeneCards. ( F , G ) The interaction between INHBA and ITGA6 was verified by Co-IF ( F ) and Co-IP ( G ) experiments. Scale bar, 50 μm. ( H ) qRT-PCR was used to detect ITGA6 mRNA expression after INHBA knockdown in NUGC-3 cells. ( I ) WB was used to detect ITGA6 protein expression after INHBA knockdown in NUGC-3 cells. ( J ) qRT-PCR was used to detect ITGA6 mRNA expression after INHBA overexpression in AGS cells. ( K ) WB was used to detect ITGA6 protein expression after INHBA overexpression in AGS cells. Data are presented as means ± SD. * p < 0.05, ** p < 0.01, **** p < 0.0001

Article Snippet: The antibodies used were as follows: GAPDH (diluted 1:500; Goodhere-Bio, Hangzhou, China; Catalog number: AB-P-R001), INHBA (diluted 1:500; Proteintech, Wuhan, China; Catalog number: 17524-1-AP), ITGA6 (diluted 1:500; Proteintech, Catalog number: 27189-1-AP), MEK1/2 (diluted 1:1000; Proteintech; Catalog number: 11049-1-AP), phospho-MEK1 (diluted 1:1000; Proteintech; Catalog number: 28930-1-AP), ERK1/2 (diluted 1:1000; Proteintech; Catalog number: 66192-1-Ig), and phospho-ERK1/2 (diluted 1:100; Proteintech; Catalog number: 28733-1-AP).

Techniques: Quantitative Proteomics, RNA Sequencing, Expressing, Co-Immunoprecipitation Assay, Quantitative RT-PCR, Knockdown, Over Expression

Journal: Oncology Research

Article Title: INHBA Promotes the Progression of Gastric Cancer by Activating MAPK Signaling Pathway via Targeting ITGA6

doi: 10.32604/or.2025.070333

Figure Lengend Snippet: Expression of ITGA6 and its interaction with INHBA in GC. ( A ) The basal expression of ITGA6 mRNA in GC cell lines and GES-1 was detected by qRT-PCR. ( B ) The basal expression of ITGA6 protein in cell lines was detected by WB. ( C ) The expression of ITGA6 protein in 30 pairs of GC paired tissues was detected by IHC. (magnification ×400). The red arrow indicates the ITGA6 expression area, which appears brown or yellow. ( D,E ) Four siRNA-ITGA6 sequences were transfected simultaneously into AGS cells. ( F,G ) The relative expression level of INHBA mRNA and protein was detected by qRT-PCR and WB after instantaneous co-transfection of INHBA siRNA and ITGA6 overexpression plasmid in NUGC-3. ( H,I ) The relative expression level of INHBA mRNA and protein were detected by qRT-PCR and WB after instantaneous co-transfection of ITGA6 siRNA and INHBA overexpression plasmid in AGS. Data are presented as means ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: The antibodies used were as follows: GAPDH (diluted 1:500; Goodhere-Bio, Hangzhou, China; Catalog number: AB-P-R001), INHBA (diluted 1:500; Proteintech, Wuhan, China; Catalog number: 17524-1-AP), ITGA6 (diluted 1:500; Proteintech, Catalog number: 27189-1-AP), MEK1/2 (diluted 1:1000; Proteintech; Catalog number: 11049-1-AP), phospho-MEK1 (diluted 1:1000; Proteintech; Catalog number: 28930-1-AP), ERK1/2 (diluted 1:1000; Proteintech; Catalog number: 66192-1-Ig), and phospho-ERK1/2 (diluted 1:100; Proteintech; Catalog number: 28733-1-AP).

Techniques: Expressing, Quantitative RT-PCR, Transfection, Cotransfection, Over Expression, Plasmid Preparation

Journal: Oncology Research

Article Title: INHBA Promotes the Progression of Gastric Cancer by Activating MAPK Signaling Pathway via Targeting ITGA6

doi: 10.32604/or.2025.070333

Figure Lengend Snippet: Oncogenic function of INHBA depends on ITGA6 and the MAPK signaling pathway. ( A ) KEGG enrichment result of the differentially expressed gene sets from the GEPIA2 database. ( B ) The expression levels of MAPK signaling pathway-related proteins were detected by WB after instantaneous co-transfection of INHBA siRNA and ITGA6 overexpression plasmid in NUGC-3. ( C,E,G,I,J ) The effects of ITGA6 on INHBA in NUGC-3 cells were detected by CCK-8 assay ( C ), colony formation assay ( E ), wound healing assay ( G ), Scale bar, 200 μm, Transwell migration assay ( I ) and Transwell invasion assay ( J ). Scale bar, 100 μm. ( D,F,H,K,L ) The effects of ITGA6 on INHBA in AGS cells were detected by CCK-8 assay ( D ), colony formation assay ( F ), wound healing assay ( H ), Scale bar, 200 μm, Transwell migration assay ( K ) and Transwell invasion assay ( L ). Scale bar, 100 μm. Data are presented as means ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: The antibodies used were as follows: GAPDH (diluted 1:500; Goodhere-Bio, Hangzhou, China; Catalog number: AB-P-R001), INHBA (diluted 1:500; Proteintech, Wuhan, China; Catalog number: 17524-1-AP), ITGA6 (diluted 1:500; Proteintech, Catalog number: 27189-1-AP), MEK1/2 (diluted 1:1000; Proteintech; Catalog number: 11049-1-AP), phospho-MEK1 (diluted 1:1000; Proteintech; Catalog number: 28930-1-AP), ERK1/2 (diluted 1:1000; Proteintech; Catalog number: 66192-1-Ig), and phospho-ERK1/2 (diluted 1:100; Proteintech; Catalog number: 28733-1-AP).

Techniques: Expressing, Cotransfection, Over Expression, Plasmid Preparation, CCK-8 Assay, Colony Assay, Wound Healing Assay, Transwell Migration Assay, Transwell Invasion Assay