Journal: Aging Cell

Article Title: Lipoteichoic acid restrains macrophage senescence via β‐catenin/ FOXO1 / REDD1 pathway in age‐related osteoporosis

doi: 10.1111/acel.14072

Figure Lengend Snippet: LTA prevents senescence‐induced FOXO1 downregulation and deactivation. Representative images (a) of double‐immunofluorescence staining of F4/80 (red) and FOXO1 (green) and quantification (b) of the percentage of FOXO1 expression area on F4/80 + cells in femoral bone. n = 6/group. Scale bars, 50 μm. BMDMs were treated with H 2 O 2 together with LTA. Representative images (c) and quantification of Western blot of the relative intensity of total FOXO1 (d) and phosphorylated FOXO1 (f). n = 3/group. Quantitative real‐time PCR analysis of FOXO1 (e) for three groups of BMDMs. n = 3/group. Representative images (g) and quantification (h) of immunofluorescence staining of nuclear FOXO1 in three groups of BMDMs. n = 3/group. Scale bars, 50 μm. Representative images (i) and quantification of Western blot of the relative intensity of cytoplasmic FOXO1 (j) and nuclear FOXO1 (k) in three groups of BMDMs. n = 3/group. * p < 0.05, ** p < 0.01, *** p < 0.001. Data are presented as mean ± SD. One‐way ANOVA with Tukey's test.

Article Snippet: For immunofluorescence staining, we incubated the cells with primary antibodies to FOXO1 (18592‐1‐AP; Proteintech, China) overnight at 4 °C, followed by incubation with Fluorescein (FITC)–conjugated Affinipure Goat Anti‐Rabbit IgG (H+L; SA00003‐2; Proteintech ) secondary antibodies.

Techniques: Double Immunofluorescence Staining, Expressing, Western Blot, Real-time Polymerase Chain Reaction, Immunofluorescence, Staining

Journal: Aging Cell

Article Title: Lipoteichoic acid restrains macrophage senescence via β‐catenin/ FOXO1 / REDD1 pathway in age‐related osteoporosis

doi: 10.1111/acel.14072

Figure Lengend Snippet: LTA inhibits mTOR phosphorylation through FOXO1/REDD1 signaling pathway. Representative images (a) of double‐immunofluorescence staining of F4/80 (red) and p‐mTOR (green) and quantification (b) of the percentage of p‐mTOR expression area on F4/80 + cells in femoral bone of vehicle‐treated 2‐month‐old mice, vehicle‐treated 12‐month‐old mice and LTA‐treated 12‐month‐old mice in the presence or absence of AS. n = 6/group. Scale bars, 50 μm. Representative images (c) of double‐immunofluorescence staining of F4/80 (red) and REDD1 (green) and quantification (d) of the percentage of REDD1 expression area on F4/80 + cells in the femoral bone of four groups of mice. n = 6/group. Scale bars, 50 μm. BMDMs were treated with H 2 O 2 together with LTA and AS. Representative images (e) and quantification of Western blot of the relative intensity of p‐mTOR (f) and REDD1 (g) in four groups of BMDMs. n = 3/group. AS: AS1842856. * p < 0.05, ** p < 0.01, *** p < 0.001. Data are presented as mean ± SD. One‐way ANOVA with Tukey's test.

Article Snippet: For immunofluorescence staining, we incubated the cells with primary antibodies to FOXO1 (18592‐1‐AP; Proteintech, China) overnight at 4 °C, followed by incubation with Fluorescein (FITC)–conjugated Affinipure Goat Anti‐Rabbit IgG (H+L; SA00003‐2; Proteintech ) secondary antibodies.

Techniques: Phospho-proteomics, Double Immunofluorescence Staining, Expressing, Western Blot

Journal: Aging Cell

Article Title: Lipoteichoic acid restrains macrophage senescence via β‐catenin/ FOXO1 / REDD1 pathway in age‐related osteoporosis

doi: 10.1111/acel.14072

Figure Lengend Snippet: LTA stimulates FOXO1 expression and promotes FOXO1 nuclear translocation through β‐catenin. Representative images (a) of double‐immunofluorescence staining of F4/80 (red) and FOXO1 (green) and quantification (b) of the percentage of FOXO1 expression area on F4/80 + cells in the femoral bone of three groups of 12‐month‐old mice. n = 6/group. Scale bars, 50 μm. BMDMs were treated with H 2 O 2 together with LTA and IWR‐1. Representative images (c) and quantification of Western blot of the relative intensity of total FOXO1 (d) and phosphorylated FOXO1 (e) in four groups of BMDMs. n = 3/group. Representative images (f) and quantification (g) of immunofluorescence staining of nuclear FOXO1 in four groups of BMDMs. n = 3/group. Scale bars, 50 μm. Representative images (h) and quantification of Western blot of the relative intensity of cytoplasmic FOXO1 (i) and nuclear FOXO1 (j) in four groups of BMDMs. n = 3/group. * p < 0.05, ** p < 0.01, *** p < 0.001. Data are presented as mean ± SD. One‐way ANOVA with Tukey's test.

Article Snippet: For immunofluorescence staining, we incubated the cells with primary antibodies to FOXO1 (18592‐1‐AP; Proteintech, China) overnight at 4 °C, followed by incubation with Fluorescein (FITC)–conjugated Affinipure Goat Anti‐Rabbit IgG (H+L; SA00003‐2; Proteintech ) secondary antibodies.

Techniques: Expressing, Translocation Assay, Double Immunofluorescence Staining, Western Blot, Immunofluorescence, Staining

Journal: Bioengineered

Article Title: Long noncoding RNA matrilineal expression gene 3 inhibits hepatocellular carcinoma progression by targeting microRNA-5195-3p and regulating the expression of forkhead box O1.

doi: 10.1080/21655979.2021.2005986

Figure Lengend Snippet: Figure 4. FOXO1 is a target gene of miR-5195-3p. (a) The binding site between miR-5195-3p and FOXO1 was predicted by bioinformatics analysis. (b) Luciferase reporter assay was used to confirm the interaction between FOXO1 and miR-5195-3p. (c) RNA pull-down assay was performed to determine whether miR-5195-3p targets FOXO1. (d, e) FOXO1 expression level was detected in miR-5195-3p overexpression or knockdown group using qPCR and Western blotting. (f) qPCR was performed to evaluate the expression of FOXO1 in HCC tissues. **P < 0.01.

Article Snippet: The The membrane was blocked with 5% nonfat milk in Tris-buffered saline with Tween 20 at room temperature for 2 h and then probed with FOXO1 and GAPDH primary antibody (1:2000 dilution; Boster, China) at 4°C overnight.

Techniques: Binding Assay, Luciferase, Reporter Assay, Pull Down Assay, Expressing, Over Expression, Knockdown, Western Blot

Journal: Bioengineered

Article Title: Long noncoding RNA matrilineal expression gene 3 inhibits hepatocellular carcinoma progression by targeting microRNA-5195-3p and regulating the expression of forkhead box O1.

doi: 10.1080/21655979.2021.2005986

Figure Lengend Snippet: Figure 5. Co-expression of MEG3 and FOXO1 inhibits HCC progression in liver cancer cell lines. (a) qPCR was performed to detect FOXO1 expression in HepG2 and Huh7 cell lines. (b) CCK-8 was used to detect HCC cell proliferation after transfection. (c, d) Transwell assay was used to assess HCC cell migration and invasion. **P < 0.01.

Article Snippet: The The membrane was blocked with 5% nonfat milk in Tris-buffered saline with Tween 20 at room temperature for 2 h and then probed with FOXO1 and GAPDH primary antibody (1:2000 dilution; Boster, China) at 4°C overnight.

Techniques: Expressing, CCK-8 Assay, Transfection, Transwell Assay, Migration

Journal: Bioengineered

Article Title: Long noncoding RNA matrilineal expression gene 3 inhibits hepatocellular carcinoma progression by targeting microRNA-5195-3p and regulating the expression of forkhead box O1.

doi: 10.1080/21655979.2021.2005986

Figure Lengend Snippet: Figure 6. Xenograft tumor model demonstrates that overexpression of MEG3 inhibites the HCC progression. (a) The images of tumor in two groups. (b) The growth curve was established. (c) Tumors weight in each group was detected. (d-f) MEG3, miR-5195-3p and FOXO1 expression level was detected in the tumor tissues using qPCR. **P < 0.01, ***P < 0.001.

Article Snippet: The The membrane was blocked with 5% nonfat milk in Tris-buffered saline with Tween 20 at room temperature for 2 h and then probed with FOXO1 and GAPDH primary antibody (1:2000 dilution; Boster, China) at 4°C overnight.

Techniques: Over Expression, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Comparison of the Chemical Components, Efficacy and Mechanisms of Action of Chrysanthemum morifolium Flower and Its Wild Relative Chrysanthemum indicum Flower against Liver-Fire Hyperactivity Syndrome of Hypertension via Integrative Analyses

doi: 10.3390/ijms232213767

Figure Lengend Snippet: Analysis of the key proteins of NF-κB and PI3K-Akt signaling pathways. ( A )—ELISA analysis of PI3K, p-PI3K, Akt, p-Akt, FOXO1, NF-κB p65, NF-κB p-p65, IκBα, respectively; ( B )—Typical bands for Western blot analysis of Akt, p-Akt, p-PI3K, NF-κB p65, and NF-κB p-p65 in C, M, CMFL, CMFH, CIFL, CIFH, P groups; ( C )—Typical bands for Western blot analysis of Akt, p-Akt, p-PI3K, NF-κB p65, and NF-κB p-p65 in C, M, CMFL, CMFM, CMFH, CIFL, CIFM, CIFH groups; ( D )—The relative expression of proteins in Western blot analysis. Values shown are means ± SD. # p < 0.05, ## p < 0.01 compared with C group; * p < 0.05, ** p < 0.01 compared with M group; ■■ p < 0.01 compared with CMFH group.

Article Snippet: Interleukin-1 beta (IL-1β, SEA563Ra), tumor necrosis factor alpha (TNF-α, SEA133Ra), interleukin-4 (IL-4, SEA077Ra), interleukin-6 (IL-6, SEA079Ra), interleukin-10 (IL-10, SEA056Ra), cyclooxygenase-2 (COX-2, SEA699Ra), angiotensin converting enzyme (ACE, SEA004Ra), and forkhead box protein O1 (FOXO1, SEA764Ra) ELISA kits were supplied by Cloud-Clone Corp. (Houston, TX, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Comparison of the Chemical Components, Efficacy and Mechanisms of Action of Chrysanthemum morifolium Flower and Its Wild Relative Chrysanthemum indicum Flower against Liver-Fire Hyperactivity Syndrome of Hypertension via Integrative Analyses

doi: 10.3390/ijms232213767

Figure Lengend Snippet: The differences in intervention effects on measurement indicators between CMF and CIF.

Article Snippet: Interleukin-1 beta (IL-1β, SEA563Ra), tumor necrosis factor alpha (TNF-α, SEA133Ra), interleukin-4 (IL-4, SEA077Ra), interleukin-6 (IL-6, SEA079Ra), interleukin-10 (IL-10, SEA056Ra), cyclooxygenase-2 (COX-2, SEA699Ra), angiotensin converting enzyme (ACE, SEA004Ra), and forkhead box protein O1 (FOXO1, SEA764Ra) ELISA kits were supplied by Cloud-Clone Corp. (Houston, TX, USA).

Techniques: