Journal:

Article Title: The classical progesterone receptor mediates Xenopus oocyte maturation through a nongenomic mechanism

doi:

Figure Lengend Snippet: Expression of xPR in Xenopus oocytes. (A) Total RNA (1 μg each) from oocytes of the various stages (stages I–III mixed with unknown ratio) were reverse-transcribed followed by PCR amplification using xPR primers encompassing exons E1 and E2. Lane 4 represents a negative control in which PCR was performed directly on input stage VI oocyte RNA without prior reverse transcription (RT). Shown is a representative of four independent experiments with arrows indicating the specifically amplified products. (B) Extracts from stage IV (lane 1), stage V (lane 2), or stage VI (lane 3) oocytes, isolated after collagenase treatment of ovarian tissues, or extracts from collagenase-treated and devitellinated oocytes (lane 4), were immunoblotted with anti-xPR. Equal amounts (50 μg) of proteins were loaded on each lane. The primary antibodies were visualized by incubation with appropriate secondary antibody-horseradish peroxidase conjugates followed by the use of a chemiluminescence kit (ECL, Amersham Pharmacia). Shown is a representative of three independent experiments. (C) Extracts from uninjected (lanes 1, 5, and 9) or Myc-xPR mRNA-injected (lanes 2, 6, and 10) oocytes (each representing half an oocyte), or extracts from untransfected (lanes 3, 7, and 11) or Myc-xPR-transfected (lanes 4, 8, and 12) COS cells (each representing one-tenth of a 3-cm dish) were immunoblotted with the indicated antibodies. Shown is a representative of three independent experiments.

Article Snippet: Antibodies against Xenopus β-integrin (8C8) were purchased from the Developmental Studies Hybridoma Bank at the University of Iowa.

Techniques: Expressing, Reverse Transcription, Amplification, Negative Control, Isolation, Incubation, Injection, Transfection

Journal:

Article Title: The classical progesterone receptor mediates Xenopus oocyte maturation through a nongenomic mechanism

doi:

Figure Lengend Snippet: xPR is a cytosolic protein in Xenopus oocytes. (A) Extracts from intact oocytes (lane 1, equivalent of half an oocyte), isolated nuclei (lane 2, 1 GV), nuclei isolated from oocytes after 15- or 45-min incubation with progesterone (lanes 3 and 4, respectively, 1 GV each), or from enucleated oocytes (lane 5, half an oocyte), were immunoblotted with anti-xPR or anti-Xenopus nucleolin (R2D2) (14). Shown is a representative of five independent experiments. (B) Extracts from intact oocytes (lane 1), 100,000 × g pellet (lane 2), or supernatant (lane 3), each of which were the equivalent of one oocyte, were blotted with anti-xPR or anti-β-integrin. Shown is a representative of five independent experiments.

Article Snippet: Antibodies against Xenopus β-integrin (8C8) were purchased from the Developmental Studies Hybridoma Bank at the University of Iowa.

Techniques: Isolation, Incubation

Journal: International journal of biological macromolecules

Article Title: Aberrant serum-derived FN1 variants bind to integrin β1 on glomerular endothelial cells contributing to thin basement membrane nephropathy.

doi: 10.1016/j.ijbiomac.2024.136282

Figure Lengend Snippet: Fig. 5. Expression changes of key GBM proteins and their response to FN1 Variants in TBMN. Immunofluorescence experiments assessed the expression of Laminin α5β2, COL4A3/4/5 and Integrin β1 in the GBM. The findings revealed non-homogeneous linear structure expression of these key GBM proteins in the presence of FN1 variants, indicating their potential involvement in the pathogenesis of TBMN. HC = healthy control.

Article Snippet: Incubating primary antibodies overnight at 4 ◦C, with specific primary antibodies against HA (1:2000, ab9110, Abcam), Integrin β1 (1:1000, 12,594–1-AP, Proteintech), β-tubulin (1:3000, ab0039, Abways), Laminin α5 (1:1000, ab210957, Abcam), Laminin β2 (1:1000, ab210956, Abcam), Laminin γ1 (1:1000, ab233389, Abcam) in Tris-Buffered Saline Tween-20 (TBST) containing 5 % skim milk.

Techniques: Expressing, Immunofluorescence, Control

Journal: International journal of biological macromolecules

Article Title: Aberrant serum-derived FN1 variants bind to integrin β1 on glomerular endothelial cells contributing to thin basement membrane nephropathy.

doi: 10.1016/j.ijbiomac.2024.136282

Figure Lengend Snippet: Fig. 6. Abnormal co-localization of FN1 variants with Integrin β1 and its impact on other GBM components. Immunofluorescence studies investigated the rela tionship between FN1 variants and the expression patterns of GBM components. The findings reveal abnormal co-localization of FN1 variants with Integrin β1, accompanied by reduced co-localization of other GBM components. *P < 0.05; **P < 0.01; ***P < 0.001.

Article Snippet: Incubating primary antibodies overnight at 4 ◦C, with specific primary antibodies against HA (1:2000, ab9110, Abcam), Integrin β1 (1:1000, 12,594–1-AP, Proteintech), β-tubulin (1:3000, ab0039, Abways), Laminin α5 (1:1000, ab210957, Abcam), Laminin β2 (1:1000, ab210956, Abcam), Laminin γ1 (1:1000, ab233389, Abcam) in Tris-Buffered Saline Tween-20 (TBST) containing 5 % skim milk.

Techniques: Immunofluorescence, Expressing

Journal: International journal of biological macromolecules

Article Title: Aberrant serum-derived FN1 variants bind to integrin β1 on glomerular endothelial cells contributing to thin basement membrane nephropathy.

doi: 10.1016/j.ijbiomac.2024.136282

Figure Lengend Snippet: Fig. 7. Competitive Binding of FN1 Variants to Integrin β1. (A and B) Duolink proximity ligation assay and Co-IP were employed to analyze the protein interaction between FN1 variants and Integrin β1. *P < 0.05; **P < 0.01; ***P < 0.001.

Article Snippet: Incubating primary antibodies overnight at 4 ◦C, with specific primary antibodies against HA (1:2000, ab9110, Abcam), Integrin β1 (1:1000, 12,594–1-AP, Proteintech), β-tubulin (1:3000, ab0039, Abways), Laminin α5 (1:1000, ab210957, Abcam), Laminin β2 (1:1000, ab210956, Abcam), Laminin γ1 (1:1000, ab233389, Abcam) in Tris-Buffered Saline Tween-20 (TBST) containing 5 % skim milk.

Techniques: Binding Assay, Proximity Ligation Assay, Co-Immunoprecipitation Assay

Journal: PLoS ONE

Article Title: miRNA-29c Suppresses Lung Cancer Cell Adhesion to Extracellular Matrix and Metastasis by Targeting Integrin β1 and Matrix Metalloproteinase2 (MMP2)

doi: 10.1371/journal.pone.0070192

Figure Lengend Snippet: (A) Effects of miR-29c on endogenous integrin β1 protein. Changes in protein expression of Int β1 was determined by Western blotting analysis in 95D 29c , 95D MiNC, 95C 29ci and 95C IhNC. GAPDH was used as an internal loading control. (B) The gray levels of Western blotting are shown by bar gragh. miR-29c mimics inhibit endogenous integrin β1 in 95D cell lines (95D 29c and 95D MiNC), while miR-29c inhibitors promote integrin β1 protein expression. ** p <0.01 (Student's t -test, n = 3). (C) Effects of miR-29c on endogenous MMP2 protein levels by Western blotting. (D) The gray levels of Western blotting are shown by bar gragh. ** p <0.01 (Student's t -test, n = 3). (E) Effects of miR-29c on MMP2 and MMP9 enzyme activity in lung cancer by gelatin zymography. (F) The gray level of every band was measured to check the differences in MMP2 enzyme activity in 95C and 95D cells (The brightest band [95D MiNC] was set to 1 unit to compare with other gray levels). ** p <0.01 (Student's t -test, n = 3). (G) The gray level of every band was measured to check the difference in MMP9 enzyme activity between 95C and 95D cells (The brightest band [95D MiNC] was set to 1 unit to compare with other gray levels). ** p <0.01 (Student's t -test, n = 3).

Article Snippet: The membranes were blocked in 5% non-fat milk in TBST buffer (Tris Buffer Saline containing 0.1% Tween-20) for 1 h at room temperature, and subsequently incubated overnight at 4°C by the appropriately diluted primary antibodies for rabbit monoclonal anti-human integrin β1 and MMP2 (diluted 1∶1000; Cell Signaling Technology).

Techniques: Expressing, Western Blot, Control, Activity Assay, Zymography

Journal: Cancer Medicine

Article Title: The expression and role of SUZ12 in lung adenocarcinoma

doi: 10.1002/cam4.70190

Figure Lengend Snippet: The effect of SUZ12 on metastasis related genes expression was tested by qRT‐PCR and western blotting. sh‐SUZ12 increased MMP1/2/3/9 mRNA expression (A) and TIMP1/2 protein expression (B), while decreased MMP14 mRNA expression (A), MMP1/9/14 (B), TIMP3 (B), and ITGB1/5(F) protein expression, without significantly effected ITGBs mRNA expression (E). oe‐SUZ12 increased MMP14 mRNA (C) and protein (D) expression, while decreased TIMP2 mRNA expression (C) and TIMP1/2 (D) protein expression.

Article Snippet: The list of primary antibodies: SUZ12 (1 μg/mL, Abcam Cambridge, cat no: ab12073), CDK2 (1:1000; Proteintech, USA, cat. no. 10122‐1‐AP), CDK3 (1:2000; Proteintech, USA, cat. no. 55103‐1‐AP), CDK6 (1:2000; Proteintech, USA, cat. no. 14052‐1‐AP), cyclin D1 (1:5000; Proteintech, USA, cat. no. 26939‐1‐AP), cyclin E1 (1:1000; Proteintech, USA, cat. no. 11554‐1‐AP), p18 (1:1000, BOSTER China, cat. no. M03299‐1), p19 (1:1000, BOSTER China, cat. no. MA1075), p53 (1:5000; Proteintech, USA, cat. no. 60283‐2‐Ig), p‐p53 (1:2000; Proteintech, USA, cat. no. 28961‐1‐AP), p57 (1:1000, BOSTER China, cat. no. BM4129), Rb (1:1000, BOSTER China, cat. no. BM4500), pRb (1:1000, BOSTER China, cat. no. BM4338), Bcl‐2 (1:1000; Proteintech, USA, cat. no. 26593‐1‐AP), Bax (1:2000; Proteintech, USA, cat. no. 50599‐2‐lg), E‐cadherin (1:5000; Proteintech, USA, cat. no. 20874‐1‐AP), N‐cadherin (1:3000; Proteintech, USA, cat. no. 22018‐1‐AP), vimentin (1:4000; Proteintech, USA, cat. no. 10366‐1‐AP), MMP1 (1:1000, BOSTER China, cat. no. A00733‐1), MMP2 (1:500, BOSTER China, cat. no. BM4075), MMP9 (1:1000, BOSTER China, cat. no. PB0709), MMP14 (1:1000, BOSTER China, cat. no. BM4119), TIMP1 (1:1000, Bioss China, cat. no. bs‐0415R), TIMP2 (1:1000, Bioss China, cat. no. bs‐10395R), TIMP3 (1:1000; Proteintech, USA, cat. no. 10858‐1‐AP), ITGB1 (1:1000, BOSTER China, cat. no. BM4308), ITGB3 (1:1000, BOSTER China, cat. no. BA1670), ITGB5 (1:1000, BOSTER China, cat. no. A04201‐1), nm23 (1:1000, BOSTER China, cat. no. BA3787), PD‐L1 (1:3000; Proteintech, USA, cat. no. 66248‐1‐Ig), and β‐actin (1:5000; Proteintech, USA, cat. no. 66009‐1‐Ig).

Techniques: Expressing, Quantitative RT-PCR, Western Blot