rabbit polyclonal klf4  (Proteintech)

Journal: iScience

Article Title: Sleep deprivation-induced sympathetic activation promotes pro-tumoral macrophage phenotype via the ADRB2/KLF4 pathway to facilitate NSCLC metastasis

doi: 10.1016/j.isci.2025.112321

Figure Lengend Snippet: KLF4 is associated with sleep deprivation-induced ADRB2 modulation of macrophage prompting on proliferation, migration, and invasion of LLC-LUC cells (A) Differentially expressed Top20 genes in metastatic lung tissues of NC group and sleep deprivation group. (B) The expression of top 5 upregulated genes in tumor tissues was analyzed by qRT-PCR. (C) KLF4 expression was detected by ADRB2-specific agonist ISO and ADRB2 antagonist ICI118,551 in macrophages treated with IL-4 in vitro . (D) The effect of ADRB2 agonist on CD206 expression in IL-4-induced KLF4-knockdown macrophages. (E) EdU assays showed the proliferation of tumor cells in the coculture system with 30% CM. CM derived from IL-4 and IL-4 + ISO induced supernatant of macrophages with KLF4 low and high expression. (F) The migration ability of tumor cells was determined by wound healing assays in the indicated coculture medium. (G) Transwell assays demonstrated invasion of tumor cells in the indicated coculture system. (H) Representative IHC images of p-JAK1 and p-STAT6 in xenograft tumors. (I) The phosphorylation of JAK1 and STAT6 expression in RAW264.7 treated with ADRB2 agonist or antagonist via Western blotting. (J) Immunoblotting of KLF4 in RAW264.7 treated with different conditions. Data represented as mean ± SD. Statistical analysis was performed using Student’s t test or Mann-Whitney U test. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001.

Article Snippet: The paraffin sections of mice tumor tissues were subjected to baking, dewaxing, dehydration, elimination of endogenous peroxidase activity, tissue pressure-resistant antigen repair, 5% BSA blocking, dropping of rabbit polyclonal antibody against KLF4 (Proteintech), rabbit polyclonal antibody against CD31 (Proteintech), rabbit polyclonal antibody against p-JAK1 (ImmunoWay), rabbit polyclonal antibody against p-STAT6 (ImmunoWay), PBS washing at 4°C overnight, incubation with secondary antibody at 37°C for 35 min, PBS washing, and staining.

Techniques: Migration, Expressing, Quantitative RT-PCR, In Vitro, Knockdown, Derivative Assay, Phospho-proteomics, Western Blot, MANN-WHITNEY

Journal: iScience

Article Title: Sleep deprivation-induced sympathetic activation promotes pro-tumoral macrophage phenotype via the ADRB2/KLF4 pathway to facilitate NSCLC metastasis

doi: 10.1016/j.isci.2025.112321

Figure Lengend Snippet: NE level is elevated in NSCLC patients with sleep deprivation and is associated with a poor prognosis (A) The concentration of NE in serum of the NSCLC patients, determined by ELISA assay. (B) Representative IHC images and quantitative analysis of ADRB2 in the NSCLC patients. (C) Representative IHC images and quantitative analysis of KLF4 in the NSCLC patients. (D) Investigation into the association between ADRB2 expression and immune cell infiltration in patients diagnosed with LUAD. (E) Analysis of the correlation between KLF4 and immune cell infiltration in LUAD patients. (F–H) Correlation analysis between ADRB2, KLF4, JAK1, and STAT6 expression. (I and J) Correlation analysis between KLF4, JAK1, and STAT6 expression. (K and L) Correlation analysis between JAK1, STAT6, and pro-tumoral macrophages. Data represented as mean ± SD. Statistical analysis was evaluated using Student’s t test. ∗∗ p < 0.01 and ∗∗∗ p < 0.001.

Article Snippet: The paraffin sections of mice tumor tissues were subjected to baking, dewaxing, dehydration, elimination of endogenous peroxidase activity, tissue pressure-resistant antigen repair, 5% BSA blocking, dropping of rabbit polyclonal antibody against KLF4 (Proteintech), rabbit polyclonal antibody against CD31 (Proteintech), rabbit polyclonal antibody against p-JAK1 (ImmunoWay), rabbit polyclonal antibody against p-STAT6 (ImmunoWay), PBS washing at 4°C overnight, incubation with secondary antibody at 37°C for 35 min, PBS washing, and staining.

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay, Expressing

Journal: iScience

Article Title: Sleep deprivation-induced sympathetic activation promotes pro-tumoral macrophage phenotype via the ADRB2/KLF4 pathway to facilitate NSCLC metastasis

doi: 10.1016/j.isci.2025.112321

Figure Lengend Snippet:

Article Snippet: The paraffin sections of mice tumor tissues were subjected to baking, dewaxing, dehydration, elimination of endogenous peroxidase activity, tissue pressure-resistant antigen repair, 5% BSA blocking, dropping of rabbit polyclonal antibody against KLF4 (Proteintech), rabbit polyclonal antibody against CD31 (Proteintech), rabbit polyclonal antibody against p-JAK1 (ImmunoWay), rabbit polyclonal antibody against p-STAT6 (ImmunoWay), PBS washing at 4°C overnight, incubation with secondary antibody at 37°C for 35 min, PBS washing, and staining.

Techniques: Recombinant, Transfection, Enzyme-linked Immunosorbent Assay, Phagocytosis Assay, Sequencing, Software

Journal: Thoracic cancer

Article Title: NSUN4 Facilitates the Activity of Oncogenic Protein CDC20 to Promote NSCLC Development by Mediating m5C Modification of CDC20 mRNA.

doi: 10.1111/1759-7714.70023

Figure Lengend Snippet: FIGURE 2 | NSUN4 inhibition affects cell growth, apoptosis, stemness, invasiveness, and migratory ability in vitro. (A–H) Various experiments were conducted using different treatment methods and cell types (A549 and SK-MES-1 NSCLC cells), with three groups: Sh-ctrl, sh-NSUN4#1, or sh-NSUN4#2. (A) Evaluation of NSUN4 protein expression by immunoblotting in cells transfected as indicated. (B) Assessment of the number of formed colonies with cells transfected as indicated. (C) Determination of cell apoptotic ratio with cells transfected as indicated by flow cytometry. (D and E) Examination of cell migratory rate and invasiveness with transfected cells. Scale bars: 100 μm. (F) Measurement of sphere formation with transfected NSCLC cells. Scale bars: 100 μm. (G and H) Expression of SOX2, CD133, and KLF4 proteins using immunoblot analysis in transfected A549 and SK-MES-1 NSCLC cells. n = 3 in (A–G). *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: We harvested protein extracts from collected tissue specimens (~50 mg) or cultivated cells (1 × 107) and conducted immunoblot analysis as described previously [18] with rabbit anti- NSUN4 polyclonal (#29786- 1- AP, 1–4000, Proteintech), rabbit antiCDC20 polyclonal (#10252- 1- AP, 1–8000, Proteintech), rabbit anti- SOX2 polyclonal (#11064- 1- AP, 1–600, Proteintech), rabbit anti- CD133 monoclonal (#ab222782, 1–2000, Abcam), rabbit anti- KLF4 polyclonal (#11880- 1- AP, 1–6000, Proteintech), or mouse anti- β- actin monoclonal (#66009- 1- Ig, 1–50 000, Proteintech).

Techniques: Inhibition, In Vitro, Expressing, Western Blot, Transfection, Flow Cytometry

Journal: Thoracic cancer

Article Title: NSUN4 Facilitates the Activity of Oncogenic Protein CDC20 to Promote NSCLC Development by Mediating m5C Modification of CDC20 mRNA.

doi: 10.1111/1759-7714.70023

Figure Lengend Snippet: FIGURE 5 | NSUN4 affects NSCLC cell malignant phenotypes through CDC20. (A–H) A549 and SK-MES-1 NSCLC cells were subjected to intro- duction with vector + sh-ctrl, NSUN4 expression construct + sh-ctrl, vector + sh-CDC20, or NSUN4 + sh-CDC20. (A) CDC20 protein expression by immunoblotting in cells transfected as indicated. (B) The number of formed colonies by colony formation assay with cells transfected as indicated. (C) Cell apoptotic ratio by flow cytometry with cells transfected as indicated. (D and E) Cell migratory rate and invasiveness by transwell assay with transfected cells. Scale bars: 100 μm. (F) Measurement of sphere formation with transfected NSCLC cells. (G and H) Expression of SOX2, CD133, and KLF4 proteins by immunoblot analysis in transfected A549 and SK-MES-1 NSCLC cells. n = 3 in (A–G). *p < 0.05, **p < 0.01, ***p < 0.001, ns: non-significant.

Article Snippet: We harvested protein extracts from collected tissue specimens (~50 mg) or cultivated cells (1 × 107) and conducted immunoblot analysis as described previously [18] with rabbit anti- NSUN4 polyclonal (#29786- 1- AP, 1–4000, Proteintech), rabbit antiCDC20 polyclonal (#10252- 1- AP, 1–8000, Proteintech), rabbit anti- SOX2 polyclonal (#11064- 1- AP, 1–600, Proteintech), rabbit anti- CD133 monoclonal (#ab222782, 1–2000, Abcam), rabbit anti- KLF4 polyclonal (#11880- 1- AP, 1–6000, Proteintech), or mouse anti- β- actin monoclonal (#66009- 1- Ig, 1–50 000, Proteintech).

Techniques: Plasmid Preparation, Expressing, Construct, Western Blot, Transfection, Colony Assay, Flow Cytometry, Transwell Assay

Journal: Thoracic cancer

Article Title: NSUN4 Facilitates the Activity of Oncogenic Protein CDC20 to Promote NSCLC Development by Mediating m5C Modification of CDC20 mRNA.

doi: 10.1111/1759-7714.70023

Figure Lengend Snippet: FIGURE 6 | NSUN4 depletion hinders the growth of A549 subcutaneous xenografts. (A–E) A549 subcutaneous xenografts were generated by implanting sh-ctrl or sh-NSUN4#2 lentivirus-infected A549 cells. After 30 days, xenografts were harvested. n = 5 for each group. (A) Growth curves of A549 subcutaneous xenografts (n = 3). (B) Representative pictures of A549 subcutaneous xenografts. (C) Tumor average weight was calculated (n = 3). (D) Expression of NSUN4, CDC20, SOX2, CD133, and KLF4 proteins by immunoblot analysis in A549 subcutaneous xenografts (n = 3). (E) Expression of NSUN4, CDC20, SOX2, CD133, and KLF4 proteins by immunohistochemistry in sections of subcutaneous xenografts. **p < 0.01, ***p < 0.001.

Article Snippet: We harvested protein extracts from collected tissue specimens (~50 mg) or cultivated cells (1 × 107) and conducted immunoblot analysis as described previously [18] with rabbit anti- NSUN4 polyclonal (#29786- 1- AP, 1–4000, Proteintech), rabbit antiCDC20 polyclonal (#10252- 1- AP, 1–8000, Proteintech), rabbit anti- SOX2 polyclonal (#11064- 1- AP, 1–600, Proteintech), rabbit anti- CD133 monoclonal (#ab222782, 1–2000, Abcam), rabbit anti- KLF4 polyclonal (#11880- 1- AP, 1–6000, Proteintech), or mouse anti- β- actin monoclonal (#66009- 1- Ig, 1–50 000, Proteintech).

Techniques: Generated, Infection, Expressing, Western Blot, Immunohistochemistry

Journal: iScience

Article Title: TRAF7 is an essential regulator of blood vessel integrity during mouse embryonic and neonatal development

doi: 10.1016/j.isci.2023.107474

Figure Lengend Snippet: Conditional deletion of Traf7 in postnatal endothelium leads to brain hemorrhage (A) Dorsal and ventral views of representative embryonic brains from P13 Traf7 iEChet and Traf7 iECko pups collected after daily tamoxifen administration. (B) Quantification of brain hemorrhagic foci in Traf7 EChet (n = 8) and Traf7 ECko (n = 8) pups. (C) H&E-stained coronal brain sections at the position indicated by the dotted blue line in (A). (D) mRNA expression analysis of Klf2 and Klf4 in brains of P7 Traf7 iEChet versus Traf7 iECko pups. Each closed symbol represents an individual embryo. Bars show the Median. Asterisks indicate significant difference: (∗) p < 0.05; (ns) not significant.

Article Snippet: Anti-KLF4, rabbit polyclonal , Sigma-Aldrich , Cat#SAB2107958.

Techniques: Staining, Expressing

Journal: iScience

Article Title: TRAF7 is an essential regulator of blood vessel integrity during mouse embryonic and neonatal development

doi: 10.1016/j.isci.2023.107474

Figure Lengend Snippet: TRAF7 and SCRIB are essential for phosphorylation of ERK5 in endothelial cells exposed to shear stress (A) Western blot analysis of ERK5 phosphorylation (p-ERK5) and TRAF7, SCRIB, MEKK3, MEKK2, KLF2, and KLF4 protein expression in HUVECs under normal (−) or shear stress (+) conditions, with addition of esiRNA specifically inhibiting each of the proteins as indicated. Anti- Renilla luciferase (RLUC) esiRNA was used as a negative control. Anti-α-Tubulin antibody was used as the loading control. The results are representative of least three independent experiments. (B) Protein gel quantification analysis of the top western blot panel in (A). p-ERK5 protein levels were calculated as percent of total ERK5 protein content in corresponding lanes using GeneTools from Syngene software. Western blots from four independent experiments were analyzed. Data are presented as Mean ± StDev.

Article Snippet: Anti-KLF4, rabbit polyclonal , Sigma-Aldrich , Cat#SAB2107958.

Techniques: Western Blot, Expressing, esiRNA, Luciferase, Negative Control, Software

Journal: iScience

Article Title: TRAF7 is an essential regulator of blood vessel integrity during mouse embryonic and neonatal development

doi: 10.1016/j.isci.2023.107474

Figure Lengend Snippet:

Article Snippet: Anti-KLF4, rabbit polyclonal , Sigma-Aldrich , Cat#SAB2107958.

Techniques: Produced, Affinity Purification, Recombinant, Transfection, Mutagenesis, Expressing, esiRNA, TaqMan Assay, Real-time Polymerase Chain Reaction, Knock-Out, Plasmid Preparation, Software