Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Long noncoding RNA AFAP1-AS1 acts as a competing endogenous RNA of miR-423-5p to facilitate nasopharyngeal carcinoma metastasis through regulating the Rho/Rac pathway

doi: 10.1186/s13046-018-0918-9

Figure Lengend Snippet: AFAP1-AS1 acts as a ceRNA to regulate FOSL2 gene expression through miR-423-5p. a . The expression of FOSL2 and its correlation with the TNM stages, lymphoma metastasis, and T stages in the same 32 NPC tissues and 13 nontumor tissues as indicated. b . The expression of FOSL2 was tightly positively correlated with that of AFAP1-AS1 in 32 NPC tissues and 13 nontumor tissues as indicated. c . The expression of FOSL2 was tightly negatively correlated with that of miR-423-5p in 32 NPC tissues and 13 nontumor tissues as indicated. d . Prediction of miR-423-5p binding sites in the FOSL2 transcript. The red nucleotides are complementary to the miR-423-5p seed sequences. e . The expression of FOSL2 was regulated by miR-423-5p at the mRNA and protein levels after transfection of miR-423-5p mimics, miR-423-5p inhibitors, or negative control in 5-8F and HNE2 cells. f . Luciferase activity was measured in 5-8F and HNE2 cells cotransfected with miR-423-5p inhibitors, miR-423-5p mimics, or negative control, and luciferase reporters containing WT or MT FOSL2 sequences, as indicated. g . The expression of FOSL2 was examined at the mRNA and protein levels in 5-8F and HNE2 cells after transfection with AFAP1-AS1 overexpression vector, or siAFAP1-AS1

Article Snippet: Membranes were incubated overnight at 4 °C with primary anti-RHOA, anti-RHOC, anti-RAC2, anti-RAB10, anti-RAB11B, anti-RAB11A, anti-RHOGDI, anti-PFN1, anti-LASP1, or anti-FOSL2 antibodies (Proteintech, Wuhan, China).

Techniques: Expressing, Binding Assay, Transfection, Negative Control, Luciferase, Activity Assay, Over Expression, Plasmid Preparation

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Long noncoding RNA AFAP1-AS1 acts as a competing endogenous RNA of miR-423-5p to facilitate nasopharyngeal carcinoma metastasis through regulating the Rho/Rac pathway

doi: 10.1186/s13046-018-0918-9

Figure Lengend Snippet: Functional analysis of FOSL2 in NPC cells. a . FOSL2 mRNA and protein levels were detected in 5-8F and HNE2 cells after transfection with the FOSL2 overexpression vector (FOSL2) or the empty vector (NC) by qRT-PCR and western blotting. b . Wound healing and transwell assays of 5-8F and HNE2 cells were conducted after transfection with pENTR- FOSL2 or pENTR empty vector. The relative ratio of wound closure per field is shown on the right. Scale bars, 100 μm. c . The expression of FOSL2 was examined at the mRNA and protein levels by qRT-PCR and western blotting in 5-8F and HNE2 cells after transfection with siFOSL2 or scrambled negative control siRNA. d . Wound healing and transwell assays were performed in 5-8F and HNE2 cells after treatment with siFOSL2 or scrambled siRNA control. The relative ratio of invasive cells per field is shown on the right. Scale bars, 100 μm. *, p < 0.05; **, p < 0.01; ***, p < 0.001

Article Snippet: Membranes were incubated overnight at 4 °C with primary anti-RHOA, anti-RHOC, anti-RAC2, anti-RAB10, anti-RAB11B, anti-RAB11A, anti-RHOGDI, anti-PFN1, anti-LASP1, or anti-FOSL2 antibodies (Proteintech, Wuhan, China).

Techniques: Functional Assay, Transfection, Over Expression, Plasmid Preparation, Quantitative RT-PCR, Western Blot, Expressing, Negative Control

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Long noncoding RNA AFAP1-AS1 acts as a competing endogenous RNA of miR-423-5p to facilitate nasopharyngeal carcinoma metastasis through regulating the Rho/Rac pathway

doi: 10.1186/s13046-018-0918-9

Figure Lengend Snippet: FOSL2 modulates the transcription of LASP1 through binding its promoter. a . Prediction of potential FOSL2 binding sites in the LASP1 promoter. The nucleotides that are consensus FOSL2 binding sequences (WT) and mutated sequences (MT) are indicated. b . The luciferase activity of the LASP1 promoter and the AP-1 transcription factor was measured in 5-8F and HNE2 cells cotransfected with the FOSL2 overexpression vector or siFOSL2 and luciferase reporters containing the AP-1 luciferase plasmid (pAP1), or the WT or MT LASP1 promoter, as indicated. Overexpression of FOSL2 significantly induced the LASP1 and AP-1 transcription activity. *p < 0.05, **p < 0.01, compared with normal control (NC) group. c . The AP-1 transcription activity was examined through transfection of the AFAP1-AS1 overexpression vector or siAFAP1-AS1 and the AP-1 reporter plasmid containing the AP-1 consensus sequence. AP-1 transcriptional activity was induced through overexpressing AFAP1-AS1 or reduced after knockdown of AFAP1-AS1 . d . The expression of LASP1 and AFAP1-AS1 was regulated by FOSL2 after knockdown or overexpression of FOSL2

Article Snippet: Membranes were incubated overnight at 4 °C with primary anti-RHOA, anti-RHOC, anti-RAC2, anti-RAB10, anti-RAB11B, anti-RAB11A, anti-RHOGDI, anti-PFN1, anti-LASP1, or anti-FOSL2 antibodies (Proteintech, Wuhan, China).

Techniques: Binding Assay, Luciferase, Activity Assay, Over Expression, Plasmid Preparation, Transfection, Sequencing, Expressing

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Long noncoding RNA AFAP1-AS1 acts as a competing endogenous RNA of miR-423-5p to facilitate nasopharyngeal carcinoma metastasis through regulating the Rho/Rac pathway

doi: 10.1186/s13046-018-0918-9

Figure Lengend Snippet: The function of AFAP1-AS1 , 423-5p, and FOSL2 in the metastasis of nasopharyngeal carcinoma 5-8F cells in vivo. a . Forty nude mice were randomly divided into 4 groups with 10 mice per group. Three experimental groups were injected with 5-8F cells transfected with the AFAP1-AS1 overexpression plasmid, miR-423-5p mimics, or the FOSL2 overexpression plasmid through the tail veins, and the control group was injected with 5-8F mock cells (NC). After 8 weeks, the nude mice were sacrificed, and the metastatic tumors were observed (upper panel). Representative lung tissue and metastatic nodes are indicated by arrows (lower panel). b . Metastatic nodes per lung in each group

Article Snippet: Membranes were incubated overnight at 4 °C with primary anti-RHOA, anti-RHOC, anti-RAC2, anti-RAB10, anti-RAB11B, anti-RAB11A, anti-RHOGDI, anti-PFN1, anti-LASP1, or anti-FOSL2 antibodies (Proteintech, Wuhan, China).

Techniques: In Vivo, Injection, Transfection, Over Expression, Plasmid Preparation

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Anticancer Action of Xiaoxianxiong Tang in Non-Small Cell Lung Cancer by Pharmacological Analysis and Experimental Validation

doi: 10.1155/2021/9930082

Figure Lengend Snippet: Information on 20 core targets.

Article Snippet: Primary antibodies against FOSL2 (cat. #: 15832-1-AP), α -tubulin (cat. #: 11224-1-AP), and GAPDH (cat. #: 10494-1-AP) were from Proteintech, and CCNA2 (cat. #: 4656) was from Cell Signaling Technology.

Techniques:

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Anticancer Action of Xiaoxianxiong Tang in Non-Small Cell Lung Cancer by Pharmacological Analysis and Experimental Validation

doi: 10.1155/2021/9930082

Figure Lengend Snippet: The cytotoxic effects of XXXT on lung cancer cells, RT-qPCR array, and western blot for core targets. (a) The inhibition rates of H460 and A549 cells treated with XXXT (0–1000 μ g/mL) for 72 h were determined using CCK8 assay. A549 or H460 treated with XXXT at IC 50 for 72 h were harvested for analysis (b–e). (b) Heatmap of 20 core targets mRNA expression in the control group and XXXT treatment group. The significant difference of core targets in A549 and H460 cells (c) was observed, including expression of CCNA2 and FOSL2 mRNA. The protein levels of CCNA2 and FOSL2 were determined by western blot (d). Statistical analysis of CCNA2 and FOSL2 proteins expression intensity (e). The above data are presented as mean ± SD for three independent experiments. NS, not significant, ∗ P < 0.05 and ∗∗ P < 0.01 showed significant difference vs. the control group.

Article Snippet: Primary antibodies against FOSL2 (cat. #: 15832-1-AP), α -tubulin (cat. #: 11224-1-AP), and GAPDH (cat. #: 10494-1-AP) were from Proteintech, and CCNA2 (cat. #: 4656) was from Cell Signaling Technology.

Techniques: Quantitative RT-PCR, Western Blot, Inhibition, CCK-8 Assay, Expressing