Journal: PLoS ONE

Article Title: Expression of Kruppel-Like Factor KLF4 in Mouse Hair Follicle Stem Cells Contributes to Cutaneous Wound Healing

doi: 10.1371/journal.pone.0039663

Figure Lengend Snippet: Primers for quantitative RT-PCR.

Article Snippet: The following primary antibodies were used: rabbit anti-KLF4 monoclonal antibody (1∶500, Genespin, Milano, Italy), rabbit anti-Ki67 antibody (1∶500, Abcam), rat anti-BrdU antibody (1∶100, AbD serotec, Oxford, UK).

Techniques:

Journal: PLoS ONE

Article Title: Expression of Kruppel-Like Factor KLF4 in Mouse Hair Follicle Stem Cells Contributes to Cutaneous Wound Healing

doi: 10.1371/journal.pone.0039663

Figure Lengend Snippet: (A to C) Quantitative RT-PCR was performed to detect the expression of KLF4, differentiation markers KLF5 and Bmp4 (A, C), and stem cells markers CD34, FEX, Cktsfb1 and DDK3 (B) in different populations isolated by FACS using anti-CD34, anti-CD49f, and anti-Lrig1 antibodies in wild type mice, a procedure that was also described in D. 1×106 sorted cells in each population were used. Values were expressed as mean ± SM of three independent experiments. * P <0.05 vs. control. (D) The proportion of CD34+/CD49f+ cells (left) purified from 6-week-old KLF4/EGFP mice was examined by flow cytometry. KLF4/EGFP-expressing cells in CD34+/CD49f+ population were shown in the right panel. Values in D were expressed as an average of three independent experiments.

Article Snippet: The following primary antibodies were used: rabbit anti-KLF4 monoclonal antibody (1∶500, Genespin, Milano, Italy), rabbit anti-Ki67 antibody (1∶500, Abcam), rat anti-BrdU antibody (1∶100, AbD serotec, Oxford, UK).

Techniques: Quantitative RT-PCR, Expressing, Isolation, Control, Purification, Flow Cytometry

Journal: PLoS ONE

Article Title: Expression of Kruppel-Like Factor KLF4 in Mouse Hair Follicle Stem Cells Contributes to Cutaneous Wound Healing

doi: 10.1371/journal.pone.0039663

Figure Lengend Snippet: (A and B) 3-day-old KLF4/EGFP mice were injected with BrdU (75mg/kg) for 5 consecutive days. BrdU-positive cells were examined 3 months later by flow cytometry (A) and immunohistochemical staining (B). Values of A were expressed as an average of three independent experiments and results of B are representative of three separate experiments. Anti-KLF4, anti-BrdU, and anti-Ki67 antibodies were used to stain consecutive slides in B. Insets show enlarged portion of the staining indicating co-localization of KLF4 and BrdU positive cells with no Ki67 signals (red arrows). Scale bars, 50 µm.

Article Snippet: The following primary antibodies were used: rabbit anti-KLF4 monoclonal antibody (1∶500, Genespin, Milano, Italy), rabbit anti-Ki67 antibody (1∶500, Abcam), rat anti-BrdU antibody (1∶100, AbD serotec, Oxford, UK).

Techniques: Injection, Flow Cytometry, Immunohistochemical staining, Staining

Journal: PLoS ONE

Article Title: Expression of Kruppel-Like Factor KLF4 in Mouse Hair Follicle Stem Cells Contributes to Cutaneous Wound Healing

doi: 10.1371/journal.pone.0039663

Figure Lengend Snippet: KLF4/CreER™/Rosa26RLacZ mice were induced by tamoxifen (100mg/kg) for 5 consecutive days at 6-week-old. 4 weeks later X-gal staining was performed. Potential KLF4 expression in interfollicular epidermis (shown by red arrows in B, C) and bulge area (A, and black arrows in B, C) was shown. A typical epithelial proliferation unit was shown in E (inset). Control staining was shown in D using KLF4/CreER™/Rosa26RLacZ mice with mock induction. Representative images were shown from 5 mice in each treatment group. Note that fixation was performed without xylene in A and B. Scale bars, 80 µm.

Article Snippet: The following primary antibodies were used: rabbit anti-KLF4 monoclonal antibody (1∶500, Genespin, Milano, Italy), rabbit anti-Ki67 antibody (1∶500, Abcam), rat anti-BrdU antibody (1∶100, AbD serotec, Oxford, UK).

Techniques: Staining, Expressing, Control

Journal: PLoS ONE

Article Title: Expression of Kruppel-Like Factor KLF4 in Mouse Hair Follicle Stem Cells Contributes to Cutaneous Wound Healing

doi: 10.1371/journal.pone.0039663

Figure Lengend Snippet: (A) Dorsal skin keratinocytes isolated from control (KLF4+/+) and KLF4 knockout (KLF4−/−) mice were analyzed by flow cytometry using mouse epidermal stem cell markers CD34 and CD49f. B. Quantification of data from A was presented. Values were expressed as mean ± SM of three independent experiments. * P <0.05 vs. control. (C) Colony formation assay was performed using total dorsal skin keratinocytes (top) or sorted CD34+/CD49f+ cells (bottom) from control (left) and KLF4 knockout (right) mice. Typical images were shown from three separate experiments. (D and E) Quantitation of the colony numbers from 2000 seeded keratinocytes in (C). Data shown were the mean ± SM of three separate experiments. * P <0.05 vs. control. Scale bars, 1 cm.

Article Snippet: The following primary antibodies were used: rabbit anti-KLF4 monoclonal antibody (1∶500, Genespin, Milano, Italy), rabbit anti-Ki67 antibody (1∶500, Abcam), rat anti-BrdU antibody (1∶100, AbD serotec, Oxford, UK).

Techniques: Isolation, Control, Knock-Out, Flow Cytometry, Colony Assay, Quantitation Assay

Journal: PLoS ONE

Article Title: Expression of Kruppel-Like Factor KLF4 in Mouse Hair Follicle Stem Cells Contributes to Cutaneous Wound Healing

doi: 10.1371/journal.pone.0039663

Figure Lengend Snippet: (A) KLF4 knockout was induced as described in . Pictures were taken 1, 5, and 10 days after two parallel 8mm wounds were placed into backs of the mice. Note that the left wounds were introduced first, and they appeared severer than the right ones because of physical stretch. (B to G) 5mm wounds were introduced into the backs of KLF4/CreER™/Rosa26RLacZ mice 5 (B, C) or 10 days (D–G) after using control (B, E) or tamoxifen (C, D, F, G) induction and X-gal staining was performed. Blue strips on epidermis were shown in C (inset 1) and G. Blue cells was indicated by black arrows outside (C) and by green arrows inside (D) the conjunction of the wound (separated by dashed green lines). Inset 2 in C showed blue cells around hair follicles. Migration of KLF4 expressing multipotent cells from hair follicles (F) and interfollicular epidermis towards the wound area was detected similarly. Results shown are representative of three independent experiments. Scale bars, 80 µm.

Article Snippet: The following primary antibodies were used: rabbit anti-KLF4 monoclonal antibody (1∶500, Genespin, Milano, Italy), rabbit anti-Ki67 antibody (1∶500, Abcam), rat anti-BrdU antibody (1∶100, AbD serotec, Oxford, UK).

Techniques: Knock-Out, Control, Staining, Migration, Expressing

Journal: PLoS ONE

Article Title: Expression of Kruppel-Like Factor KLF4 in Mouse Hair Follicle Stem Cells Contributes to Cutaneous Wound Healing

doi: 10.1371/journal.pone.0039663

Figure Lengend Snippet: (A) KLF4 expression was examined in control (siCon) and KLF4 knockdown (siKLF4) cells by real time PCR. * P <0.05 vs. control. (B) Down regulation of KLF4 was accompanied by reduced expression of its downstream target p21 by Western Blotting analysis (Left) and the corresponding densitometry quantification (Right). (C) Flow cytometric analysis of cancer stem cells using CD44 and podoplanin as the markers in siCon and siKLF4 cells. Values were expressed as an average of three independent experiments. (D) Scratch assays were performed using siCon and siKLF4 cells. 2×10 6 cells were cultured to confluence, then scratched and photographed immediately (0 h) or after 12 hours (12 h). (E) Quantification of cell migration during 12 h after scratching. Error bars represent standard error from three separate experiments. * P <0.05 vs. control.

Article Snippet: The following primary antibodies were used: rabbit anti-KLF4 monoclonal antibody (1∶500, Genespin, Milano, Italy), rabbit anti-Ki67 antibody (1∶500, Abcam), rat anti-BrdU antibody (1∶100, AbD serotec, Oxford, UK).

Techniques: Expressing, Control, Knockdown, Real-time Polymerase Chain Reaction, Western Blot, Cell Culture, Migration

Journal: FASEB BioAdvances

Article Title: Inhibition of the Hippo pathway by verteporfin reduces the proliferation and stemness of rat hair follicle neural crest stem cells under hypoxia

doi: 10.1096/fba.2025-00025

Figure Lengend Snippet: Antibodies information.

Article Snippet: Mouse monoclonal anti‐KLF4 , Santa Cruz Biotechnology , Cat# sc‐393,462.

Techniques:

Journal:

Article Title: Kr?ppel-Like Factor-4 Transcriptionally Regulates VE-cadherin Expression and Endothelial Barrier Function

doi: 10.1161/CIRCRESAHA.110.219592

Figure Lengend Snippet: A-F, Untreated or ECs treated with LiCl (20 ng/mL) or with recombinant WNT3A (50 ng/mL) for 3 d and stained for VE-cadherin. Representative images of control and treated ECs at 200× (A, C, E) and 400× (B, D, F) magnification. For additional images, see Figure S2. G-J) Cell extracts were analyzed by Western blot (WB) for VE-cadherin, β-catenin, KLF4, and GAPDH. Results are representative of at least three separate experiments.

Article Snippet: Mouse anti-human KLF4 mAb (H00009314-M01) was purchased from AbNOVA (Walnut, CA).

Techniques: Recombinant, Staining, Control, Western Blot

Journal:

Article Title: Kr?ppel-Like Factor-4 Transcriptionally Regulates VE-cadherin Expression and Endothelial Barrier Function

doi: 10.1161/CIRCRESAHA.110.219592

Figure Lengend Snippet: A, Timeline of TER assay. B, HLMVECs plated on gold microelectrodes were left untreated (control) or transfected with a non-silencing siRNA or KLF4-silencing siRNA, followed by TER assay. Note that KLF4 silencing inhibited the thrombin response relative to untreated group (no transfection) or negative control group (non-silencing siRNA; n = 5 per group). Mean value (± s.e.m.) of maximal TER responses to thrombin (50 nM) stimulation (n = 7). Thrombin-induced decrease in TER was significantly attenuated in HLMVECs transfected by KLF4-depletion compared with untreated control or negative control group transfected with a non-silencing siRNA. C-E, KLF4 knockdown increases transendothelial permeability of fluorescein isothiocyanate (FITC)-conjugated albumin by decreasing VE-cadherin expression and AJ integrity. C, Timeline of experiments. D, Confluent HLMVEC monolayers were grown on microporous filters for 36 h, either left alone (control) or treated with control siRNA or with KLF4-siRNA for 12 hours. At 18 hr post transfection, transendothelial FITC-albumin permeability was measured. Control HLMVECs showed basal transendothelial FITC-albumin permeability values, while KLF4 knockdown increased transendothelial FITC-albumin permeability. Re-expression of VE-cadherin into KLF4-depleted ECs partially restored the effect of loss of KLF4. Values are mean ± s.e.m. (n= 10). *p < 0.05 vs other control (untreated) group. **p <0.01 KLF4 siRNA vs control siRNA. E, The efficiency of KLF4-knockdown and VE-cadherin re-expression in HLMVECs was determined by Western blotting.

Article Snippet: Mouse anti-human KLF4 mAb (H00009314-M01) was purchased from AbNOVA (Walnut, CA).

Techniques: Control, Transfection, Negative Control, Knockdown, Permeability, Expressing, Western Blot

Journal:

Article Title: Kr?ppel-Like Factor-4 Transcriptionally Regulates VE-cadherin Expression and Endothelial Barrier Function

doi: 10.1161/CIRCRESAHA.110.219592

Figure Lengend Snippet: A, Schematic of human VE-cadherin promoter −1.3 kb upstream of transcription start site (TSS). Potential KLF4 binding (CACCC) sites are indicated. B, Sequence of human VE-cadherin primer pair used for ChIP experiments. C, HLMVECs and HUVECs were grown in complete media, and left untreated (-) or treated for 3 days with WNT3A. ChIP assay was performed with indicated antibodies. PCR product of VE-cadherin promoter using input chromatin. D, Biotin-labeled oligonucleotide probes (P1-P4) containing CACCC sites used for EMSA. E, Representative image of EMSA blot. Probe-1 (P1) of KLF4 site from VE-cadherin-promoter was incubated with nuclear extracts prepared from ECs treated with WNT3A or by pre-incubation of nuclear extract with anti-KLF4 antibody in the presence or absence of cold-unlabeled oligonucleotide. Results are representative of at least three separate experiments.

Article Snippet: Mouse anti-human KLF4 mAb (H00009314-M01) was purchased from AbNOVA (Walnut, CA).

Techniques: Binding Assay, Sequencing, Labeling, Incubation

Journal:

Article Title: Kr?ppel-Like Factor-4 Transcriptionally Regulates VE-cadherin Expression and Endothelial Barrier Function

doi: 10.1161/CIRCRESAHA.110.219592

Figure Lengend Snippet: A, Time-line of siRNA administration, LPS challenge, myeloperoxidase (MPO) as a measure of lung neutrophil sequestration, and lung extravascular water content assays. B, Lung MPO activities were assayed in mice receiving either control siRNA or Klf4-specific siRNA with or without LPS challenge at 20 min, 1 h, 3 h, and 6 h. C, Lung tissue extracts were prepared 18 h after siRNA administration (but prior to receiving LPS) and efficacy of Klf4-knockdown in lung tissues was evaluated by immunoblotting with the indicated antibodies. Values are mean ± s.e.m. (n=12 per group). *p < 0.01 vs control (untreated) group. **p <0.05 Klf4 siRNA vs control siRNA.

Article Snippet: Mouse anti-human KLF4 mAb (H00009314-M01) was purchased from AbNOVA (Walnut, CA).

Techniques: Control, Knockdown, Western Blot

Journal: Investigative Ophthalmology & Visual Science

Article Title: MYEOV Is a Novel Marker of Differentiated Corneal Epithelium

doi: 10.1167/iovs.66.14.7

Figure Lengend Snippet: Regulation of MYEOV expression by PAX6 and KLF4 transcription factors. ( A ) Comparative analyses of PAX6 , KLF4 , MYEOV , and KRT12 mRNA expression in the setting of PAX6 KD using five distinct siRNAs and KLF4 KD using three distinct siRNAs in cultured human corneal epithelial cells ( n = 4 donors for PAX6 KD, n = 6 donors for KLF4 KD). Error bars represent SD. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. ( B ) Representative western blot analysis of PAX6, KLF4, MYEOV, KRT12, and β-actin (loading control) protein expression by PAX6 KD ( n = 3 donors) and KLF4 KD ( n = 5 donors).

Article Snippet: Primary antibodies used were rabbit anti-β-actin pAb (1:1000, 5125; Cell Signaling Technology), rabbit anti-MYEOV pAb (1:1000, MBS9611913; MyBioSource, San Diego, CA, USA), rabbit anti-KRT12 mAb (1:5000, ab185627; Abcam), rabbit anti-PAX6 mAb (1:1000, ab195045; Abcam), and mouse anti-KLF4 mAb (1:500, sc-166238; Santa Cruz Biotechnology).

Techniques: Expressing, Cell Culture, Western Blot, Control

Journal: Investigative Ophthalmology & Visual Science

Article Title: MYEOV Is a Novel Marker of Differentiated Corneal Epithelium

doi: 10.1167/iovs.66.14.7

Figure Lengend Snippet: KLF4 binding sites within promoter/enhancer regions of MYEOV . KLF4 binding sites were mapped using data from JASPAR 2024 and ReMap 2022 Atlas using the UCSC genome browser. ENCODE candidate cis -regulatory elements (cCREs) show the regions of promoter-like signature ( red ), proximal enhancer-like signature ( orange ), and distal enhancer-like signature ( yellow ) of MYEOV.

Article Snippet: Primary antibodies used were rabbit anti-β-actin pAb (1:1000, 5125; Cell Signaling Technology), rabbit anti-MYEOV pAb (1:1000, MBS9611913; MyBioSource, San Diego, CA, USA), rabbit anti-KRT12 mAb (1:5000, ab185627; Abcam), rabbit anti-PAX6 mAb (1:1000, ab195045; Abcam), and mouse anti-KLF4 mAb (1:500, sc-166238; Santa Cruz Biotechnology).

Techniques: Binding Assay

Journal: Oncotarget

Article Title: Transcriptional repression of FOXO1 by KLF4 contributes to glioma progression

doi: 10.18632/oncotarget.13184

Figure Lengend Snippet: A. Schematic of the putative binding site of KLF4 in the FOXO1 promoter. B. KLF4 mRNA level in glioma tissue samples represented as fold change were detected with qRT-PCR by normalizing to GAPDH as endogenous control and the expression level in matched non-tumor tissues was set as 1. C. The correlation between KLF4 mRNA expression and glioma grades was analyzed. D. Pearson's correlation analyses between relative KLF4 mRNA and FOXO1 mRNA levels in glioma tissues. E. Representative images of KLF4 protein level in glioma tissue samples detected by IHC (20×). F. The negative correlation between KLF4 protein and FOXO1 protein level in glioma tissues was analyzed. G. Kaplan-Meier curves showing the overall survival of patients with high or low protein level of KLF4 in their gliomas. Statistical significance was determined by a log-rank test.

Article Snippet: The primary antibodies used for analysis included rabbit anti-FOXO1 monoclonal antibody, rabbit anti-KLF4 monoclonal antibody and mouse anti-β-Actin monoclonal antibody were from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Binding Assay, Quantitative RT-PCR, Control, Expressing

Journal: Oncotarget

Article Title: Transcriptional repression of FOXO1 by KLF4 contributes to glioma progression

doi: 10.18632/oncotarget.13184

Figure Lengend Snippet: A. FOXO1 expression in mRNA and protein levels was detected by qRT-PCR and western blot after KLF4 knockdown. vs control, * p <0.0001. B. ChIP-qPCR for the KLF4 binding to the FOXO1 promoter in KLF4 knockdowned cell lines by transfection of shRNAs. vs control, * p <0.001. C. Luciferase reporter assay for the luciferase activity driven by FOXO1 promoter in U87 or U251 cell lines after KLF4 knockdown. vs control, * p <0.001.

Article Snippet: The primary antibodies used for analysis included rabbit anti-FOXO1 monoclonal antibody, rabbit anti-KLF4 monoclonal antibody and mouse anti-β-Actin monoclonal antibody were from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Knockdown, Control, ChIP-qPCR, Binding Assay, Transfection, Luciferase, Reporter Assay, Activity Assay

Journal: Oncotarget

Article Title: Transcriptional repression of FOXO1 by KLF4 contributes to glioma progression

doi: 10.18632/oncotarget.13184

Figure Lengend Snippet: A. KLF4 knockdown decreased invasion potential of U87 and U251 cell lines, as shown by cell counts per area. B. KLF4 knockdown inhibited proliferation potential of U87 and U251 cell lines in vitro determined cell number count. C. KLF4 knockdown inhibited U87 and U251 cells growth in vivo. Data are represented as a mean ± SD from three mice. vs control, * p <0.001.

Article Snippet: The primary antibodies used for analysis included rabbit anti-FOXO1 monoclonal antibody, rabbit anti-KLF4 monoclonal antibody and mouse anti-β-Actin monoclonal antibody were from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Knockdown, In Vitro, In Vivo, Control