Fig 2: Expression of AMHR2 protein (green) in human oocytes, as revealed by confocal microscopy. A Horizontal section of an AMHR2-positive immature oocytes. B Immature oocyte positively stained for AMHR2 protein. C Horizontal section of an AMHR2-positive mature oocyte. D Mature oocyte positively stained for AMHR2 protein. Images were taken with a confocal microscope at 630 × magnification. AMHR2 is stained green, and genetic material is stained blue (DAPI)

Fig 3: Differential gene expression analysis of The Cancer Genome Atlas (TCGA) RNA sequence data. (a) Volcano plot showing differential gene expression between the AMHR2‐high and AMHR2‐low groups among TCGA lung adenocarcinoma cases. Genes represented above the horizontal line are significantly differentially expressed with an FDR‐adjusted p‐value <0.01. Red dots represent genes with a fold‐change increase >1.2 in the AMHR2‐low group, indicating upregulation, while blue dots represent genes with a fold‐change decrease >1.2 in the AMHR2‐low group, indicating downregulation. (b) Pathway enrichment analysis of the genes significantly upregulated in the AMHR2‐low group. Results from both KEGG (upper section) and biological process (lower section) enrichment are shown. (c) Gene set enrichment analysis of RNA‐sequencing data from TCGA lung adenocarcinoma cases revealed an enrichment of cell cycle‐related genes in the AMHR2‐low group compared to the AMHR2‐high group. (d) Violin plots showing the expression of representative genes that respectively promote the cell cycle. Each dot is one sample. Red represents the AMHR2‐high group, and blue represents the AMHR2‐low group. Horizontal bars are presented as the 25th percentile (bottom quartile), median, and 75th percentile (top quartile). p‐values were obtained using the Mann–Whitney U test (a p‐value <0.05 was significant).

Fig 4: Effects of recombinant human AMH treatment on cell proliferation and cell cycle related gene expression in non‐small cell lung cancer cell lines overexpressing AMHR2. (a) Inhibitory effect of recombinant human AMH (rhAMH) on the proliferation of AMHR2‐overexpressing cell lines. Cell counts were obtained after 5 days of treatment with 20 or 50 ng/mL rhAMH. The y‐axis is normalized with control group mean values set to 100. Data from one representative of three independent experiments are shown, with error bars representing standard deviation. Statistical analysis was performed using Welch's t‐test. (*p < 0.05; **p < 0.01; ***p < 0.001). (b) Investigation of the effects of rhAMH on the genes using qPCR. The EGFP‐induced control A549 cell line (EGFP: left bar) as the control, AMHR2‐overexpressing A549 cell line (AMHR2: middle bar) and AMHR2‐overexpressing A549 cell line treated with 50 mg/mL rhAMH (AMHR2 + rhAMH: right bar) were compared. The qPCR was performed in triplicate, and data from one representative of six independent experiments are shown. The y‐axis is normalized with control group mean values set to 100, with error bars representing standard deviation. Statistical analysis was performed using Welch's t‐test. (*p < 0.05; **p < 0.01; ***p < 0.001).

Fig 5: Immunohistochemical detection of AMHR2 in non‐small cell lung cancer and ovarian serous adenocarcinoma tissues. (a) Immunostaining of ovarian serous adenocarcinoma tissue. (b) Normal lung tissue. (c) Representative lung adenocarcinoma tissue showing positive staining for AMHR2. (d) Lung adenocarcinoma tissue showing negative staining for AMHR2. (e) Lung squamous cell carcinoma tissue. All micrographs were captured at 400× magnification.