Journal: Frontiers in Cell and Developmental Biology

Article Title: Pharmacological inhibition of FOXO1 promotes lymphatic valve growth in a congenital lymphedema mouse model

doi: 10.3389/fcell.2022.1024628

Figure Lengend Snippet: Western blot antibodies.

Article Snippet: KLF4 , NBP217070 (Novus Biologicals) , 1:333.

Techniques: Western Blot, Diagnostic Assay

Journal: PLoS ONE

Article Title: Cancer stem cell subpopulations in primary colon adenocarcinoma

doi: 10.1371/journal.pone.0221963

Figure Lengend Snippet: Representative 3,3-diaminobenzidine immunohistochemical-stained images showing protein expression of induced-pluripotent stem cell markers OCT4 (A-C, brown), SOX2 (D-F, brown), NANOG (G-I, brown), KLF4 (J-L, brown) and c-MYC (M-O, brown) in normal colon (A,D,G,J,M), low-grade (B,F,H,K,N) and high-grade (C,F,I,L,O) colon adenocarcinoma tissue samples. Nuclei were counter-stained with hematoxylin (A-O, blue). Original magnification: 400x.

Article Snippet: Staining was carried out on the Leica BOND TM RX Auto-stainer using primary antibodies for OCT4 (1:30; cat#MRQ-10, Cell Marque, Rocklin, CA, USA), SOX2 (1:200; cat#ab97959, Abcam, Cambridge, MA, USA), NANOG (1:200; cat#EP225, Cell Marque), KLF4 (1:200; cat#NBP2-24749SS, Novus Biologicals LLC, Littleton, CO, USA) and c-MYC (1:1000; cat#ab32, Abcam).

Techniques: Immunohistochemical staining, Staining, Expressing

Journal: PLoS ONE

Article Title: Cancer stem cell subpopulations in primary colon adenocarcinoma

doi: 10.1371/journal.pone.0221963

Figure Lengend Snippet: Percentage of cell population stained positively for induced-pluripotent stem cell markers OCT4, SOX2, NANOG, KLF4 and c-MYC by 3,3-diaminobenzidine immunohistochemical staining, for the epithelium (A) and the stroma (B). Normal colon samples from patients with low-grade colon adenocarcinoma (LGCA; pale blue, n = 9) are displayed separately to normal colon samples from patients with high-grade colon adenocarcinoma (HGCA; dark blue, n = 8). LGCA samples are shown in pale yellow (n = 10), and HGCA samples are shown in dark yellow (n = 8). Statistical significance with a p -value between 0.05 and 0.01 is shown by *, and that for <0.01 is represented by **. Error bars show standard error.

Article Snippet: Staining was carried out on the Leica BOND TM RX Auto-stainer using primary antibodies for OCT4 (1:30; cat#MRQ-10, Cell Marque, Rocklin, CA, USA), SOX2 (1:200; cat#ab97959, Abcam, Cambridge, MA, USA), NANOG (1:200; cat#EP225, Cell Marque), KLF4 (1:200; cat#NBP2-24749SS, Novus Biologicals LLC, Littleton, CO, USA) and c-MYC (1:1000; cat#ab32, Abcam).

Techniques: Staining, Immunohistochemical staining

Journal: PLoS ONE

Article Title: Cancer stem cell subpopulations in primary colon adenocarcinoma

doi: 10.1371/journal.pone.0221963

Figure Lengend Snippet: Representative immunofluorescence immunohistochemical-stained images showing protein expression of induced-pluripotent stem cell markers OCT4 (A-J, green), KLF4 (A-C, red), NANOG (D-G, H-J, red), SOX2 (H-J, red), and c-MYC (H-J, green) in normal colon (A,D,E,H), low-grade (B,E,F,I) and high-grade (C,F,G,J) colon adenocarcinoma tissue samples. Cell nuclei were counter-stained with 4’, 6’-diamidino-2-phenylindole (A-L, blue). Original magnification: 400x.

Article Snippet: Staining was carried out on the Leica BOND TM RX Auto-stainer using primary antibodies for OCT4 (1:30; cat#MRQ-10, Cell Marque, Rocklin, CA, USA), SOX2 (1:200; cat#ab97959, Abcam, Cambridge, MA, USA), NANOG (1:200; cat#EP225, Cell Marque), KLF4 (1:200; cat#NBP2-24749SS, Novus Biologicals LLC, Littleton, CO, USA) and c-MYC (1:1000; cat#ab32, Abcam).

Techniques: Immunofluorescence, Immunohistochemical staining, Staining, Expressing

Journal: PLoS ONE

Article Title: Cancer stem cell subpopulations in primary colon adenocarcinoma

doi: 10.1371/journal.pone.0221963

Figure Lengend Snippet: mRNA expression of induced-pluripotent stem cell markers OCT4 (A), SOX2 (B), NANOG (C), KLF4 (D) and c-MYC (E) detected by RT-qPCR. Data displayed as the fold-change of gene expression in tumor samples relative to their patient-matched normal colon sample (Y-axis). A cohort of six LGCA tissue samples are shown in blue, and a cohort of six HGCA tissue samples are shown in yellow (X-axis).

Article Snippet: Staining was carried out on the Leica BOND TM RX Auto-stainer using primary antibodies for OCT4 (1:30; cat#MRQ-10, Cell Marque, Rocklin, CA, USA), SOX2 (1:200; cat#ab97959, Abcam, Cambridge, MA, USA), NANOG (1:200; cat#EP225, Cell Marque), KLF4 (1:200; cat#NBP2-24749SS, Novus Biologicals LLC, Littleton, CO, USA) and c-MYC (1:1000; cat#ab32, Abcam).

Techniques: Expressing, Quantitative RT-PCR, Gene Expression

Journal: PLoS ONE

Article Title: Cancer stem cell subpopulations in primary colon adenocarcinoma

doi: 10.1371/journal.pone.0221963

Figure Lengend Snippet: Representative images of in-situ hybridization, showing mRNA expression of iPSC genes OCT4 (A-C, brown), SOX2 (D-F, brown), NANOG (G-I, brown), KLF4 (J-L, brown) and c-MYC (M-O, brown) in the epithelial cells ( arrows ) and stromal cells ( arrowheads ) in normal colon (A,D,G,J,M), low-grade (B,E,H,K,N) and high-grade (C,F,I,L,O) colon adenocarcinoma tissue samples. Nuclei were counter-stained with hematoxylin (blue). Original magnification: 1000x.

Article Snippet: Staining was carried out on the Leica BOND TM RX Auto-stainer using primary antibodies for OCT4 (1:30; cat#MRQ-10, Cell Marque, Rocklin, CA, USA), SOX2 (1:200; cat#ab97959, Abcam, Cambridge, MA, USA), NANOG (1:200; cat#EP225, Cell Marque), KLF4 (1:200; cat#NBP2-24749SS, Novus Biologicals LLC, Littleton, CO, USA) and c-MYC (1:1000; cat#ab32, Abcam).

Techniques: In Situ Hybridization, Expressing, Staining

Journal: International Journal of Biological Sciences

Article Title: The KLF4/Galectin-3 cascade is a key determinant of tubular cell death and acute kidney injury

doi: 10.7150/ijbs.110790

Figure Lengend Snippet: The Galectin-3 promoter is bound and transactivated by KLF4. (A) Heatmap analysis of transcription factor activity predicted by the SCENIC package for proximal tubular cells across the indicated groups. (B) The potential promoter sequences of Galectin-3 bound by the transcription factor KLF4, as predicted by the JASPAR database. (C) Chromatin immunoprecipitation-polymerase chain reaction (ChIP-PCR) assays showing PCR amplification of Galectin-3 chromatin corresponding to the region of the promoter (site1: nucleotides -1991 to -2000 and site2: -1443 to -1452 presented in (B)) immunoprecipitated with anti-KLF4 or with control IgG antibody from cisplatin-challenged kidneys. (D) Western blot and graphic presentation showing changes in KLF4 expression in HK2 cells treated with cisplatin (25 μg/ml) or H 2 O 2 (500 mM) at different time points as indicated. (E) Western blot for KLF4 and Galectin-3 protein in HK2 cells transfected with scramble or KLF4 siRNA and followed by cisplatin (25 μg/ml) for 12 hours. One of the three independent experiments with identical results was shown. (F) Representative immunofluorescence staining images for Galectin-3 in HK2 cells transfected with scramble or KLF4 siRNA. Scale bar = 25 μm. (G) KLF4 protein expression in HK2 cells transfected with KLF4-OE plasmid. One of the three independent experiments with identical results was shown. (H) Galectin-3 protein levels in HK2 cells transfected with empty vector control or KLF4-OE plasmid followed by cisplatin treatment. (I) Semiquantitative analysis of Galectin-3 protein from (H) (n = 3). (J) Schematic illustration of Galectin-3 promoter reporter constructs containing the wild-type KLF4 binding sequences (BS WT) and the corresponding mutant sequences (BS Mut) used in luciferase assays. (K) Relative activation of WT and mutant Galectin-3 promoter by KLF4 in 293T cells. The luciferase activity of each group was normalized to that co-transfected with pECMV-NC and pgl4 plasmid (n = 4). Data are presented as means ± SEM. * p ˂0.05 or *** p ˂0.001.

Article Snippet: Mice with a floxed KLF4 allele were ordered from Cyagen (cat: S-CKO-03285, Guangzhou, China, C57BL/6J background).

Techniques: Activity Assay, Chromatin Immunoprecipitation, Polymerase Chain Reaction, Amplification, Immunoprecipitation, Control, Western Blot, Expressing, Transfection, Immunofluorescence, Staining, Plasmid Preparation, Construct, Binding Assay, Mutagenesis, Luciferase, Activation Assay

Journal: International Journal of Biological Sciences

Article Title: The KLF4/Galectin-3 cascade is a key determinant of tubular cell death and acute kidney injury

doi: 10.7150/ijbs.110790

Figure Lengend Snippet: Blocking the KLF4/Galectin-3 signaling cascade attenuates kidney tubular cell death. (A) Western blot for cleaved PARP and cleaved caspase 3 protein in HK2 cells transfected with scramble or KLF4 siRNA followed by cisplatin exposure. (B) Semiquantitative analysis for cleaved PARP and cleaved caspase 3 protein from (A) (n = 3). (C) Representative flow cytometry plots analyzing KLF4 and scramble siRNA- treated HK2 cells, which were then treated with either vehicle or cisplatin. (D) Summary data quantifying apoptosis among different groups in (C) (n = 3). (E-F) Propidium Iodide (PI) staining assay (E) and quantitative analysis (F) of PI in HK2 cells among groups indicated. Scale bar = 50 μm. (G) Western blot for cleaved PARP and cleaved caspase 3 in HK2 cells with pECMV-KLF4 and/ or Galectin-3 siRNA followed by cisplatin treatment. (H) Semiquantitative analysis for cleaved PARP and cleaved caspase 3 protein levels from (G) (n = 3). (I) KLF4 and Galectin-3 protein expression in HK2 cells pretreated with Kenpaullone and followed by cisplatin (25 μg/ml) for 12 hrs. One of the three independent experiments with identical results was shown. (J) Western blot showing cleaved PARP and cleaved caspase 3 protein levels in HK2 cells pretreated with different concentrations of Kenpaullone followed by cisplatin exposure. (K) Representative flow cytometry plots and quantitative analyses of Kenpaullone- and vehicle-treated HK2 cells, as indicated by the groups (n = 3). (L) PI staining assay and quantitative analysis of PI in HK2 cells among groups indicated (n=3). Scale bar = 50 μm. (M) Western blots showing KLF4 and Galectin-3 protein expression in HK2 cells for the indicated group. One of the three independent experiments with identical results was shown. (N) Western blot analysis showing the levels of cleaved PARP and cleaved caspase 3 proteins in HK2 cells pretreated with various concentrations of APTO-253 before exposure to cisplatin. (O-P) PI staining assay (O) and quantitative analysis (P) of PI in HK2 cells among indicated groups (n=3). Scale bar = 50 μm. Ken, Kenpaullone; PI, Propidium Iodide. Data are presented as means ± SEM. * p ˂0.05, ** p ˂0.01, or *** p ˂0.001.

Article Snippet: Mice with a floxed KLF4 allele were ordered from Cyagen (cat: S-CKO-03285, Guangzhou, China, C57BL/6J background).

Techniques: Blocking Assay, Western Blot, Transfection, Flow Cytometry, Staining, Expressing

Journal: International Journal of Biological Sciences

Article Title: The KLF4/Galectin-3 cascade is a key determinant of tubular cell death and acute kidney injury

doi: 10.7150/ijbs.110790

Figure Lengend Snippet: KLF4 protein is induced in tubular cells from patients and mouse models with acute kidney injury, and deletion of KLF4 in proximal tubular cells attenuates cisplatin-induced AKI. (A) Representative immunohistochemical staining images showing the expression of KLF4 in kidney tubular cells from patients with acute kidney injury (AKI). Red arrows indicating the KLF4 positive tubular cells. Scale bar = 50 μm. (B) Representative staining images showing colocalization of KLF4 and Galectin-3 proteins in kidney sections from patients with AKI. White arrow heads indicating double positive tubular cells. Scale bar = 50 μm. (C-D) Western blot assay (C) and semiquantitative analysis (D) showing the abundance of KLF4 protein in the mouse kidneys after cisplatin exposure at day 2 and 3 (n = 3). (E) Linear regression analysis of KLF4 and Galectin-3 expression levels in the kidneys of cisplatin mouse model. (F-G) Western blot assay (F) and semiquantitative analysis (G) showing the expression of KLF4 in the kidneys after IRI at day 1 and 3 (n = 3). (H) Linear regression analysis of KLF4 and Galectin-3 expression levels in the kidneys of IRI mouse model. (I) Strategy for generating mice with kidney proximal tubular-specific deletion of KLF4. (J) Genotyping the mice by PCR analysis of genomic DNA. (K) Representative immunofluorescence staining for KLF4 protein in WT and PKO kidney sections after cisplatin treatment. Scale bar = 50 μm. (L-O) KLF4 and Galectin-3 mRNA (L-M) and protein (N-O) expression levels in kidneys from WT and PKO mice following cisplatin exposure (n = 5). (P) Kidney histology from the groups as shown by PAS staining and kidney pathology scores (n = 5). Scale bar =100 μm. (Q) Serum creatinine and BUN among groups as indicated (n = 5). AKI, acute kidney injury; IRI, ischemia-reperfusion injury. Data are presented as means ± SEM. * p ˂0.05, ** p ˂0.01, or *** p ˂0.001.

Article Snippet: Mice with a floxed KLF4 allele were ordered from Cyagen (cat: S-CKO-03285, Guangzhou, China, C57BL/6J background).

Techniques: Immunohistochemical staining, Staining, Expressing, Western Blot, Immunofluorescence

Journal: International Journal of Biological Sciences

Article Title: The KLF4/Galectin-3 cascade is a key determinant of tubular cell death and acute kidney injury

doi: 10.7150/ijbs.110790

Figure Lengend Snippet: Tubular KLF4 deficiency attenuates kidney injury, apoptosis and inflammatory response. (A) Principal component analysis of global transcriptomics from WT and PKO kidneys following cisplatin challenge. (B) Heatmap of significant gene expression from WT and PKO kidneys with cisplatin exposure. (C) Renal mRNA expression levels of KIM-1, NGAL and Hnf4a in cisplatin-exposed kidneys from WT and PKO mice (n = 5). (D) Western blots for KIM-1, NGAL and cleaved caspase 3 in kidneys from WT and MKO mice after cisplatin injection at day 3. (E) Semiquantitative determination of protein abundance in (D) (n = 5). (F) Representative kidneys stained with KIM-1 and NGAL protein. Scale bar = 50 μm. (G) Representative images and quantification of TUNEL staining in kidney sections from WT and PKO mice with cisplatin nephropathy (n = 5). Scale bar = 50 μm. (H) Representative immunofluorescence staining for F4/80 and Ly6G in cisplatin-exposed kidneys from different groups as indicated. Scale bar = 50 μm. (I) Quantitative analysis for F4/80-positive macrophages and Ly6G-positive neutrophils in cisplatin-exposed kidneys among groups as indicated (n = 5). (J) The IL6, TNFa, and MCP-1 mRNA expression levels in WT and PKO kidneys following cisplatin treatment (n = 5). Data are presented as means ± SEM. * p ˂0.05, ** p ˂0.01, or *** p ˂0.001.

Article Snippet: Mice with a floxed KLF4 allele were ordered from Cyagen (cat: S-CKO-03285, Guangzhou, China, C57BL/6J background).

Techniques: Gene Expression, Expressing, Western Blot, Injection, Quantitative Proteomics, Staining, TUNEL Assay, Immunofluorescence

Journal: International Journal of Biological Sciences

Article Title: The KLF4/Galectin-3 cascade is a key determinant of tubular cell death and acute kidney injury

doi: 10.7150/ijbs.110790

Figure Lengend Snippet: KLF4 deletion in proximal tubular cells ameliorates IRI-induced kidney injury and inflammatory response. (A) Representative images for PAS staining in kidneys among groups as indicated. Scale bar = 100 μm. (B) Kidney pathology scores in (A) (n = 5). (C) Serum creatinine and BUN levels in WT and PKO mice following IRI (n = 5). (D) Renal mRNA levels for KLF4 and Galectin-3 in IRI model (n = 5). (E) Western blot assay and semiquantitative analysis for Galectin-3 protein in IRI kidneys from different groups as indicated (n = 5). (F) Representative immunohistochemical staining for Galectin-3 protein in IRI kidneys among groups as indicated. Scale bar = 50 μm. (G-H) Western blot assay (G) and semiquantitative analysis (H) for KIM-1, NGAL and cleaved caspase 3 protein in IRI kidneys (n = 5). (I) Representative immunohistochemical staining for KIM-1 and NGAL protein in IRI kidneys among groups as indicated. Scale bar = 50 μm. (J-K) Representative images (J) and quantitative analysis (K) of TUNEL staining in the indicated groups (n = 5). Scale bar = 50 μm. (L-M) Representative immunochemical staining (L) and quantitative analysis of F4/80 and Ly6G (M) in IRI-induced kidneys from the indicated groups (n = 5). Scale bar = 50 μm. (N) Renal mRNA expression levels for IL6, TNFa and MCP-1 in IRI kidneys among groups as indicated (n = 5). Data are presented as means ± SEM. * p ˂0.05, ** p ˂0.01, or *** p ˂0.001.

Article Snippet: Mice with a floxed KLF4 allele were ordered from Cyagen (cat: S-CKO-03285, Guangzhou, China, C57BL/6J background).

Techniques: Staining, Western Blot, Immunohistochemical staining, TUNEL Assay, Expressing

Journal: International Journal of Biological Sciences

Article Title: The KLF4/Galectin-3 cascade is a key determinant of tubular cell death and acute kidney injury

doi: 10.7150/ijbs.110790

Figure Lengend Snippet: Inhibition of KLF4 signaling with Kenpaullone attenuates cisplatin-induced acute kidney injury. (A-B) Western blot assay (A) and semiquantitative analysis (B) for KLF4 and Galectin-3 protein in cisplatin-exposed kidneys among groups as indicated (n = 6). (C) Representative images for PAS staining in cisplatin-exposed kidneys and kidney pathology scores (n = 6). Scale bar = 100 μm. (D) Serum creatinine and BUN levels in groups as indicated (n = 6). (E-F) Western blot assay (E) and semiquantitative analysis (F) for KIM-1, NGAL and cleaved caspase 3 protein in cisplatin-exposed kidneys among groups as indicated (n = 6). (G) Representative images for KIM-1 and NGAL staining in cisplatin-exposed kidneys from vehicle and Kenpaullone-treated mice. Scale bar = 50 μm. (H-I) TUNEL staining (H) and quantification analysis (I) of kidney sections from vehicle and Kenpaullone-treated mice following cisplatin exposure (n = 6). Scale bar = 50 μm. (J-K) Representative immunofluorescence staining (J) and quantitative analysis (K) of F4/80 and Ly6G in cisplatin-treated kidneys from the indicated groups (n = 5). Scale bar = 50 μm. (L) IL-6, TNFa, and MCP-1 mRNA abundance in vehicle and Ken-treated kidneys following cisplatin exposure (n = 6). Data are presented as means ± SEM. * p ˂0.05, ** p ˂0.01, or *** p ˂0.001.

Article Snippet: Mice with a floxed KLF4 allele were ordered from Cyagen (cat: S-CKO-03285, Guangzhou, China, C57BL/6J background).

Techniques: Inhibition, Western Blot, Staining, TUNEL Assay, Immunofluorescence

Journal: bioRxiv

Article Title: Circular RNA Circ-Cdr1as modulates Macrophage phenotype and Cardiac Reparative Function by Circ-Cdr1as-miR-7-Klf4 pathway

doi: 10.1101/2025.02.21.639391

Figure Lengend Snippet: (A) qRT-qPCR of KLF4 expression in 3D post-MI left ventricular (LV) tissue compared to sham, normalized to 18S. N=3/group. (Unpaired T-test) (B) RT-qPCR of KLF4 expression in 3D post-MI left ventricular tissue in AAV9-GFP and AAV9-circ-cdr1as groups, normalized to 18S. N=4 (Unpaired-t-test). (C) Changes in KLF4 expression in naïve, pro-inflammatory, and anti-inflammatory macrophages overexpressing circ-cdr1as or knockdown of circ-cdr1as, normalized to 18S. N= 3/group (One-way ANOVA). (D) Changes in KLF4 expression in naïve, pro-inflammatory, and anti-inflammatory macrophages treated with miR-7b-5p mimic or anti-mir-7b-5p, normalized to miR-24. N= 3/group (One-way ANOVA). (E) FACS analysis of F4/80/CD86 + cells (pro-inflammatory MΦ marker) or (F) F4/80/CD206+ cells (anti-inflammatory MΦ marker) in naïve macrophages treated with lentivirus KFL4 or shRNA KLF4 and their respective controls. N=3-4/group (One-way ANOVA). Data are mean ± SEM. NS, non-significant, *p<0.05, ** p<0.01, *** p<0.001. MΦ, macrophages; pro-inflammatory marker: CD86; anti-inflammatory marker: CD206.

Article Snippet: For overexpression of Klf4, BMDMs were transfected with Klf4 (NM_010637) Mouse Tagged ORF Clone Lentiviral Particle (Origene) (titer concentration 1.7×10^7 TU/mL).

Techniques: Expressing, Quantitative RT-PCR, Knockdown, Marker, shRNA

Journal: Frontiers in Physiology

Article Title: KLF4-Induced Connexin40 Expression Contributes to Arterial Endothelial Quiescence

doi: 10.3389/fphys.2019.00080

Figure Lengend Snippet: Cx40 expression is gradually regulated by shear stress. (A) KLF4 expression in bEnd.3 cells under static conditions (St) or exposed to 24 h of LLSS and HLSS was assessed by qPCR. N = 3. (B) Cx40 expression in bEnd.3 cells under static conditions (St) or exposed to 24 h of LLSS and HLSS was assessed by qPCR. N = 3. (C) Representative images of Cx40 expression (green) in bEnd.3 cells under static conditions (St) or exposed to LLSS and HLSS for 24 h. Arrow indicates the direction of flow. Nuclei were stained with DAPI (blue). Scale bar represents 10 μm. (D) Schematic representation of flow regions in cell cultures induced by orbital rotation; green = HLSS and red = OSS. (E) Cx40 expression in bEnd.3 cells under static conditions (St) or exposed to 48 h of HLSS was assessed by Western blotting. N = 5. (F) Phase-contrast images of HUVECs under static conditions or exposed for 48 h of HLSS. (G,H) Cx40 expression in HUVECs under static conditions (St) or exposed to 48 h of HLSS was assessed by Western blotting (G) or qPCR ( H ; N = 6).

Article Snippet: Ten micrograms of digested, cross-linked chromatin was immunoprecipitated using 2 μg anti-KLF4 antibody (R&D systems) and protein magnetic beads.

Techniques: Expressing, Shear, Staining, Western Blot

Journal: Frontiers in Physiology

Article Title: KLF4-Induced Connexin40 Expression Contributes to Arterial Endothelial Quiescence

doi: 10.3389/fphys.2019.00080

Figure Lengend Snippet: KLF4 regulates flow-dependent Cx40 expression. KLF4 (A) , Cx40 (B) , KLF2 (C) , Cx37 (D) and Cx43 (E) expression in bEnd.3 cells transfected with KLF4 siRNA or NT siRNA and subsequently exposed to 48 h of HLSS or kept under static conditions. N = 6. (F) Representative ChIP results for bEnd.3 chromatin precipitated with antibodies against KLF4. Three different CACCC KLF-consensus sites (indicated in bp) were amplified for Cx40 (left) and one for FSP1 (right; positive control). Levels of DNA are normalized to input. N = 2.

Article Snippet: Ten micrograms of digested, cross-linked chromatin was immunoprecipitated using 2 μg anti-KLF4 antibody (R&D systems) and protein magnetic beads.

Techniques: Expressing, Transfection, Amplification, Positive Control