The primary antibodies used in the study: pPI3K110 obtained from Bioss Antibodies (USA), PTEN, pPTEN, P70S6, Cyclin B1, pAkt, Akt1, pmTOR and mTOR from Cell Signaling (Beverly, USA), FoxF2, PI3K110, Cyclin D1, ß-actin, p27 and p-p27 from Abcam (Cambridge, UK), and p21 and p-p21 from Santa-Cruz (USA). The secondary antibodies: Alexafluor 633 obtained from Life Technologies, anti-rabbit HRP-conjugate from Biorad (USA) and anti-mouse HRP-conjugate from GE Heath Care (Buckinghamshire, UK).
Foxf2
Manufactured by Abcam
Sourced in United States
FOXF2 is a transcription factor that plays a role in regulating gene expression. It is involved in various cellular processes, including development, differentiation, and homeostasis. FOXF2 contains a forkhead DNA-binding domain and is known to bind to specific DNA sequences to activate or repress target genes.
Automatically generated - may contain errors
Lab products found in correlation
Cyclin b1, by Cell Signaling Technology (1 mentions)
P pten, by Cell Signaling Technology (1 mentions)
Anti rabbit hrp conjugate, by Bio-Rad (1 mentions)
P akt, by Cell Signaling Technology (1 mentions)
Alexa fluor 633, by Thermo Fisher Scientific (1 mentions)
Cyclin d1, by Abcam (1 mentions)
P mtor, by Cell Signaling Technology (1 mentions)
P smad3, by Abcam (1 mentions)
Rpe65, by Novus Biologicals (1 mentions)
Ssea4, by Abcam (1 mentions)
Triton x 100, by Merck Group (1 mentions)
α sma, by Abcam (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
4 6 diamidino 2 phenylindole dihydrochloride dapi, by Merck Group (1 mentions)
P smad2, by Abcam (1 mentions)
Rhodopsin, by Abcam (1 mentions)
Nanog, by Proteintech (1 mentions)
Tyrp1, by Abcam (1 mentions)
C myc, by Proteintech (1 mentions)
Flag tag (flag), by MBL Life Science (1 mentions)
5 protocols using foxf2
1
Signaling Pathway Protein Analysis
2
Immunofluorescence Analysis of Stem Cell Markers
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For immunofluorescence analysis, fixed cells, cryosections from eyes and matrigel containing transplanted cells in nude mice, were permeabilized with 0.25% Triton X-100 (Sigma) for 2 min, washed with PBS, and then blocked with 2% bovine serum albumin (BSA, Sigma) in PBS. The sections were incubated with the primary antibodies (1:200) against OCT4, NANOG, SOX2, ZO-1, FN1, LIN7A, PARD6B, CRALBP, BEST1, PPM1A, MITF-A, CRX and C-MYC (Proteintech, Rosemont, IL); against SSEA4, TYRP1, α-SMA, FOXF2, MYOSIN, p-SMAD2, p-SMAD3, NR2E1, Rhodopsin, and Ki67 (Abcam, Cambridge, UK); against RPE65 (Novus Biologicals, Centennial, CO), and against FLAG (1:1000, MBL International, Woburn, MD) overnight at 4°C. They were then washed three times with PBS, followed by incubation with the fluorescent secondary antibodies (1:1000, invirogen) overnight. F-actin was stained with phalloidin-iFluor 555 (Yeasen Biotech, Shanghai, China). 4,6 diamidino-2-phenylindole dihydrochloride (DAPI, Sigma) was used to indicate the nucleus. The samples were then examined by fluorescence microscope (Olympus IX73, Tokyo, Japan). Antibodies were listed in key resources table .
3
Foxf2 and Smad6 Interaction in hESCs
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HESCs were harvested after stimulation of TGF‐β1 for 72 hours. Catch and Release® v2.0 Reversible Immunoprecipitation System (Millipore, USA) was used for Co‐IP test. The procedure followed the manufacturer's instruction. The cells were lysed, and then, 2 μg/mL of antibodies (Foxf2 or Smad6) was used to precipitate proteins. Anti‐rabbit immunoglobulin G monoclonal antibody (1:1000, Sigma‐Aldrich) was used as negative control in the experiments. The precipitated proteins were resolved to SDS‐PAGE and transferred to a PVDF membrane. The membranes were incubated with primary antibodies against Foxf2 or Smad6 (Abcam) overnight at 4°C, followed with secondary anti‐rabbit horseradish peroxidase‐conjugated IgG (1:1000, CST, USA), developed in ChemiDoc™ XRS+Imaging System using the chemiluminescence method (ECL, Millipore).
4
Protein Expression Analysis in Tumor Tissues
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Tumor tissues and adjacent tissues which were ground in liquid nitrogen were collected. hEEC and Hela cells of each group were also collected. RIPA lysis buffer was used to extract total proteins in these tissues and cells. In addition, nucleus proteins of Hela cells of each group were also obtained through using nucleus protein extraction kit (Boster Biological Technology, Ltd., Wuhan, China). Separation of proteins was conducted by SDS/PAGE at 120 V. Then 2 h blocking with skimmed milk (5%) was performed at room temperature. Primary antibodies used in the present study were rabbit anti-mouse FOXF2, E-cadherin, Vimentin, Snail, β-catenin, c-Myc, CyclinDl, MMP9, and Lgr5, respectively (1:1000, Abcam, U.S.A.). After 12-h incubation with primary antibody at 4°C, three-times washing by TBST were implemented. Subsequently, goat anti-rabbit IgG secondary antibody (1:2000, Beijing Zhongshan Jinqiao Biotechnology Co., Ltd., China) was added to incubate for 1 h at room temperature. TBST was also used for washing three times. GAPDH was set as internal reference of total proteins in cells. Nucleus proteins of Hela cells were normalized to Histone H3.
5
Western Blot Analysis of FOXF2 Protein
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The protocols for western blotting were described in a previous study (Zhang et al., 2019 (link)). The primary antibodies were incubated at 4°C overnight according to the following dilution ratio: FOXF2 (1:200; Abcam, Cambridge, MA, United States). β-actin and Histone H3 were used as endogenous loading control.
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