Journal: Reproductive and Developmental Medicine

Article Title: Elevated levels of KLF12 impair trophoblast syncytialization via GCM1 downregulation

doi: 10.1097/rd9.0000000000000099

Figure Lengend Snippet: Fig. 1. KLF12 expression in placental villi from women with miscarriage vs normal pregnancies. (A) Gestational weeks of women who had normal pregnancies (n = 10) and those who experienced miscarriage (n = 11). (B) KLF12 immunodetection in placental villi from women with normal pregnancies and miscarriages. GAPDH served as the internal control. (C) Scatter plots showing the KLF12 expression data for all placental villi obtained from women with normal pregnancies and miscarriages. ***P <0.001. (D) Immunohistochemical analysis of KLF12 in placental villi obtained from women who had normal pregnancies and miscar- riages. Scale bar represents 100 μm. GAPDH: glyceraldehyde-3-phosphate dehydrogenase; KLF12: Kruppel-like factor 12.

Article Snippet: The primary antibodies against KLF12 (Santa Cruz Biotechnology, Cat. No. sc84347, dilution 1:200), cytokeratin (Abcam, Cat. No. ab9377, dilution 1:200), MCT1 (Millipore, Cat. No. AB1286, dilution 1:300), and MCT4 (Millipore, Cat. No. AB3314P, dilution 1:200) were used for this purpose.

Techniques: Expressing, Immunodetection, Control, Immunohistochemical staining

Journal: Reproductive and Developmental Medicine

Article Title: Elevated levels of KLF12 impair trophoblast syncytialization via GCM1 downregulation

doi: 10.1097/rd9.0000000000000099

Figure Lengend Snippet: Fig. 2. KLF12 levels during BeWo cell syncytialization. KLF12 and hCG immunodetection in BeWo cells treated with either DMSO or FSK for 24 and 48 h. GAPDH served as the internal control. DMSO: dimethylsulfoxide; FSK: forskolin; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; hCG: human chorionic gonadotropin; KLF12: Kruppel-like factor 12.

Article Snippet: The primary antibodies against KLF12 (Santa Cruz Biotechnology, Cat. No. sc84347, dilution 1:200), cytokeratin (Abcam, Cat. No. ab9377, dilution 1:200), MCT1 (Millipore, Cat. No. AB1286, dilution 1:300), and MCT4 (Millipore, Cat. No. AB3314P, dilution 1:200) were used for this purpose.

Techniques: Immunodetection, Control

Journal: Reproductive and Developmental Medicine

Article Title: Elevated levels of KLF12 impair trophoblast syncytialization via GCM1 downregulation

doi: 10.1097/rd9.0000000000000099

Figure Lengend Snippet: Fig. 3. Effects of elevated KLF12 levels on BeWo cell syncytialization. (A) hCG immunodetection in BeWo cells transfected with Ad-GFP-KLF12 or control and stimulated with FSK for 24 and 48 h. GAPDH served as the internal control. (B) Immunofluorescence analysis of E-Cadherin in BeWo cells transfected with Ad-GFP-KLF12 or control and stimulated with FSK for 48 h. The white dotted line marks the syncytia. Scale bar represents 100 μm. (C) Histological and immunohistochemical analysis of cytokeratin, MCT4, and MCT1 in WT and KLF12 OE placentas at embryonic day 9.5. The black dotted line marks the trophoblast. Scale bars represent 200 μm (HE staining), 500 μm (cytokeratin staining), and 100 μm (MCT4 and MCT1 staining). FSK: forskolin; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; hCG: human chorionic gonadotropin; HE: hematoxylin-eosin; KLF12: Kruppel-like factor 12; KLF12 OE: KLF12 overexpression; WT: wild-type.

Article Snippet: The primary antibodies against KLF12 (Santa Cruz Biotechnology, Cat. No. sc84347, dilution 1:200), cytokeratin (Abcam, Cat. No. ab9377, dilution 1:200), MCT1 (Millipore, Cat. No. AB1286, dilution 1:300), and MCT4 (Millipore, Cat. No. AB3314P, dilution 1:200) were used for this purpose.

Techniques: Immunodetection, Transfection, Control, Immunofluorescence, Immunohistochemical staining, Staining, Over Expression

Journal: Reproductive and Developmental Medicine

Article Title: Elevated levels of KLF12 impair trophoblast syncytialization via GCM1 downregulation

doi: 10.1097/rd9.0000000000000099

Figure Lengend Snippet: Fig. 4. KLF12 regulation of GCM1 expression. (A) Quantitative real-time PCR analysis of syncytialization-related genes in BeWo cells transfected with Ad-GFP- KLF12 or control and stimulated with FSK for 24 h. **P <0.01; ***P <0.001. (B) Chromatin immunoprecipitation–PCR using primers against the human GCM1 promoter region (top). PCR products were separated by agarose gel electrophoresis (below). (C) Relative GCM1-Luc activity in BeWo cells transfected with Flag- KLF12 or control and stimulated with FSK for 24 h. *P <0.05. (D) Diagram illustrating the regulatory hierarchy between KLF12 overexpression and pregnancy outcomes. DMSO: dimethylsulfoxide; FSK: forskolin; GCM-1: glial cells missing-1; hCG: human chorionic gonadotropin; KLF12: Kruppel-like factor 12; PCR: polymerase chain reaction; TSS: transcription start site.

Article Snippet: The primary antibodies against KLF12 (Santa Cruz Biotechnology, Cat. No. sc84347, dilution 1:200), cytokeratin (Abcam, Cat. No. ab9377, dilution 1:200), MCT1 (Millipore, Cat. No. AB1286, dilution 1:300), and MCT4 (Millipore, Cat. No. AB3314P, dilution 1:200) were used for this purpose.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Transfection, Control, Chromatin Immunoprecipitation, Agarose Gel Electrophoresis, Activity Assay, Over Expression, Polymerase Chain Reaction

Journal: Translational Oncology

Article Title: Krüppel-like factor 12 decreases progestin sensitivity in endometrial cancer by inhibiting the progesterone receptor signaling pathway

doi: 10.1016/j.tranon.2024.102041

Figure Lengend Snippet: KLF12 expression negatively correlated with PGR in EC tissues. (A) IHC to detect the expression of KLF12 and PGR in normal endometrial and EC tissues. (B and C) Statistical charts of KLF12 and PGR expression in EC tissues at each stage. *P<0.05, **P<0.01, ***P<0.001.

Article Snippet: Lentiviruses encoding GFP-KLF12 (KLF12) or GFP (as control) were purchased from Keygen.

Techniques: Expressing

Journal: Translational Oncology

Article Title: Krüppel-like factor 12 decreases progestin sensitivity in endometrial cancer by inhibiting the progesterone receptor signaling pathway

doi: 10.1016/j.tranon.2024.102041

Figure Lengend Snippet: KLF12 expression negatively correlated with PGR in EC cell lines. (A) Relative expression of KLF12 protein in four EC cell lines (Ishikawa, HEC-1B, AN3CA, MFE-296). (B and C) Relative expression of KLF12 protein and mRNA after overexpression of KLF12 in Ishikawa cells. (D and E) Relative expression of KLF12 protein and mRNA after stable knockdown of KLF12 in MFE296 cells. (F and G) Western blotting and qPCR detected protein and mRNA expression, respectively, of PGR in Ishikawa cells overexpressing KLF12. (H and I) Western blotting and qPCR detected PGR protein and mRNA expression, respectively, after KLF12 knockdown in MFE296 cells. *P<0.05, **P<0.01, ***P<0.001, ns: not statistically significant.

Article Snippet: Lentiviruses encoding GFP-KLF12 (KLF12) or GFP (as control) were purchased from Keygen.

Techniques: Expressing, Over Expression, Knockdown, Western Blot

Journal: Translational Oncology

Article Title: Krüppel-like factor 12 decreases progestin sensitivity in endometrial cancer by inhibiting the progesterone receptor signaling pathway

doi: 10.1016/j.tranon.2024.102041

Figure Lengend Snippet: The relationship of KLF12 with progesterone sensitivity. (A and B) CCK-8 assay to detect drug resistance and calculate IC50. (C and G) Cell proliferation was measured by CCK-8 assay after treatment with 2 μm MPA. (E and H) Clone formation assay to determine the proliferative activity and clonal formation ability after treatment with 2 μm MPA. (E I) Cell apoptosis was detected by flow cytometry. (F and J) The proportion of cells in different cycle phases, as observed with flow cytometry. *P<0.05, **P<0.01, ***P<0.001, ns: Not statistically significant.

Article Snippet: Lentiviruses encoding GFP-KLF12 (KLF12) or GFP (as control) were purchased from Keygen.

Techniques: CCK-8 Assay, Tube Formation Assay, Activity Assay, Flow Cytometry

Journal: Translational Oncology

Article Title: Krüppel-like factor 12 decreases progestin sensitivity in endometrial cancer by inhibiting the progesterone receptor signaling pathway

doi: 10.1016/j.tranon.2024.102041

Figure Lengend Snippet: KLF12 regulates the promoter region of PGR. (A) Gene number captured by Ish-ovklf12 and Ish-vector. (B and C) DNA fragments bound to KLF12 captured by Ish-ovklf12 and Ish-vector. (D) IGV visualization software was used to analyze the amount of DNA captured; the higher the peak, the more the quantity of DNA captured. (E) The binding site of the PGR promoter region and KLF12. (F) Luciferase assays were performed after transfection with pGL3-basic-PGR mut and pGL3-basic-PGR wt . Relative luciferase activity was analyzed after 48 h treatment. (G) The main pathway for target gene enrichment that binds to KLF12. *P<0.05, **P<0.01, ***P<0.001.

Article Snippet: Lentiviruses encoding GFP-KLF12 (KLF12) or GFP (as control) were purchased from Keygen.

Techniques: Plasmid Preparation, Software, Binding Assay, Luciferase, Transfection, Activity Assay

Journal: Translational Oncology

Article Title: Krüppel-like factor 12 decreases progestin sensitivity in endometrial cancer by inhibiting the progesterone receptor signaling pathway

doi: 10.1016/j.tranon.2024.102041

Figure Lengend Snippet: KLF12 affects progesterone sensitivity via PGR. (A and B) Relative expression of KLF12 and PGR proteins and mRNAs after PGR knockdown in MFE296 cells and (C and D) 296-shklf12 cells. (E and G) Cell proliferation was compared by CCK-8 and (F and H) clone formation experiments. (I) Cell apoptosis was detected by flow cytometry. (J) Proportion of cells in different cycle phases, as detected by flow cytometry. *P<0.05, **P<0.01, ***P<0.001.

Article Snippet: Lentiviruses encoding GFP-KLF12 (KLF12) or GFP (as control) were purchased from Keygen.

Techniques: Expressing, Knockdown, CCK-8 Assay, Flow Cytometry

Journal: Translational Oncology

Article Title: Krüppel-like factor 12 decreases progestin sensitivity in endometrial cancer by inhibiting the progesterone receptor signaling pathway

doi: 10.1016/j.tranon.2024.102041

Figure Lengend Snippet: KLF12 overexpression decreases EC progesterone sensitivity in vivo . (A) Images of subcutaneous neoplasia at day 42. Ish-ovklf12 and Ish-vector cells were treated with MPA. (B) The subcutaneous tumor volume was measured at different time points, and the growth trend was noted. (C) Immunohistochemistry of mouse tumor tissues for KLF12 and PGR expression detection in Ish-ovklf12 + MPA and Ish-vector + MPA groups. Ip: intraperitoneal injection, *P<0.05, **P<0.01, ****P<0.001, ns: not statistically significant.

Article Snippet: Lentiviruses encoding GFP-KLF12 (KLF12) or GFP (as control) were purchased from Keygen.

Techniques: Over Expression, In Vivo, Plasmid Preparation, Immunohistochemistry, Expressing, Injection