Fig 1: Kruppel-Like Factor 4 (KLF4) and Specificity protein-1 (SP1) contribute to IPO13 dependent oxidative stress induced cell death.IPO13+/+ ESCs were treated without or with siRNAs targeting SP1 or KLF4. 3 d post transfection, cells were treated with H2O2 as indicated for 1 h, and then stained either immediately (“No recovery”) or subsequent to 2 h recovery as indicated, for propidium iodide (PI) and flow cytometric analysis. a Representative dot plots of the indicated samples with the percentage of ESC PI positive. b Pooled data (IPO13+/+ 125 µM (n = 7), IPO13+/+ 300–600 µM (n = 9), IPO13-/- 125–600 µM (n = 9), IPO13+/+ +KLF4 siRNA 125–600 µM (n = 7), IPO13+/+ + SP1 siRNA 125–600 µM (n = 7)) for % PI positive cells relative to untreated (UT) control from the analysis such as that in a are shown (mean ± SEM), where p values (two tailed student’s t test) top to bottom left to right are no recovery: p = 0.6160, p = 0.0032, p = 0.0210, p = 0.0215, p = 0.0198, p = 0.0029, p = 0.0048, p = 0.0326, p = 0.0074, and 2 h recovery: p = 0.6160, p = 0.3882, p = 0.0273, p = 0.6300, p = 0.6484, p = 0.0123, p = 0.1919, p = 0.3598, p = 0.0278. Source data are provided as a Source Data file.
Fig 2: Silencing of lncRNA XIST inhibits angiogenesis and increases cerebral vascular injury following CIS in vivo(A) Images show the IF or dual-IF staining for Itga5, KLF4, CD31 with Ki67, three CAMs (E-selectin, VCAM-1, and ICAM-1), Claudin-5, and ZO-1 in the ischemic hemisphere taken from sham or the negative control shRNA (si-Ctl) and shRNA-lncRNA XIST (si-XIST)-treated mice at day 7 post-ischemic reperfusion. Scale bar, 100 µm (insets, 30 µm). (B) Quantification of Itga5, KLF4, CD31/Ki67, E-selectin, VCAM-1, ICAM-1, Claudin-5, and ZO-1 expressions. Results are expressed as the mean ± standard deviation and analyzed by one-way ANOVA (n = 6 mice per experimental group). Note that compared to si-Ctl-treated mice, the number of Itga5-, KLF4-, Claudin-5-, and ZO-1-positive events and CD31/Ki67 dual-positive cells was all significantly reduced, but the number of three CAMs, including E-selectin-, VCAM-1-, and ICAM-1-positive events, was all remarkably induced in the si-XIST-treated mice at day 7 post-ischemic reperfusion. ##p < 0.01 and ###p < 0.001 versus control (sham); *p < 0.05 and **p < 0.01 versus si-Ctl-treated mice.
Fig 3: Increased expression of three cell adhesion molecules and KLF4 following focal cerebral ischemia. a Images show IF staining for three cell adhesion molecules (E-selectin, VCAM-1, and ICAM-1) and KLF4 in ischemic hemisphere from sham-operated mice (S, control) or mice at day 1, 2, 4, 7, and 14 post-ischemia. Scale bar = 100 µm. b Quantification of E-selectin, VCAM-1 and ICAM-1, and KLF4 expressions. Results are expressed as the mean ± standard deviation of the number of positive events per field of view (n = 6 per experimental group). Note that cerebral ischemia induced a strong increase in the expression of all the three cell adhesion molecules in both ischemic penumbra and core, reaching a peak at day 2 and then declining at day 4. While in the ischemic penumbra, the number of KLF4-positive events increased slightly during the first 2 days post-ischemia, but then increased significantly by day 4, and reached a maximum between 7 and 14 days post-ischemia. However, in the ischemic core, the number of KLF4-positive events increased markedly during the first 2 days post-ischemia, before declining at day 4. *P < 0.05, **P < 0.01, ***P < 0.001 compared with control
Fig 4: KLF4 expression is diminished in fibrotic fibroblasts.Baseline KLF4 expression in fibroblasts outgrown from lungs of patients with IPF and control nonfibrotic (normal) lungs by real-time quantitative PCR (qPCR) analysis (A) and protein densitometry of Western blots (B). (C) Correlation analysis between the baseline KLF4 and FOXM1 mRNA expression in normal (blue dots) and IPF (red dots) fibroblasts; the cell lines displayed in C are an unselected subset of those displayed in A. Baseline KLF4 expression in fibroblasts outgrown from mouse lungs on day 21 after bleomycin and saline by qPCR analysis (D) and protein densitometry of Western blots (E). KLF4 expression in CCL210 cells after 48 hours of TGF-ß (2 ng/mL) stimulation analyzed by qPCR (F) and Western blot (G). Expression of KLF4 protein in CCL210 cells after 3 hours of PGE2 (0.5 M) treatment analyzed by Western blot (results depicted from 1 experiment representative of 3 experiments (H). In A–C, each symbol represents a single patient lung-derived fibroblast line, and in D and E, each symbol represents an individual murine lung-derived fibroblast line. In A, B, and D–F, mRNA and protein values are normalized to GAPDH and expressed relative to the control (normal human or saline-treated mouse). Data in A–F are shown as mean ± SEM. *P < 0.05, 2-way ANOVA.
Fig 5: SNHG14 regulated KLF4 expression by acting as a miR-214-3p sponge. (A,B) SK-N-SH cells were transfected with si-NC, si-SNHG14, si-SNHG14 + anti-miR-NC, or si-SNHG14 + anti-miR-214-3p before MPP+ (1 mM, 24 h) stimulation, and then KLF4 mRNA and protein levels were gauged by qRT-PCR and western blot assays. *P < 0.05. (C) Schematic model of the SNHG14/miR-214-3p/KLF4 axis in MPP+-stimulated cytotoxicity in PD. MPP+ stimulation elevated the expression of SNHG14 in SK-N-SH cells. Then, the elevated expression of SNHG14 downregulated miR-214-3p expression, and thus upregulated KLF4 level in MPP+-stimulated SK-N-SH cells. Finally, the overexpression of KLF4 accelerated MPP+-stimulated cytotoxicity in PD.
Supplier Page from Abcam for Anti-KLF4 antibody