Journal: Oncotarget

Article Title: Adipose-derived mesenchymal stem cells promote osteosarcoma proliferation and metastasis by activating the STAT3 pathway

doi: 10.18632/oncotarget.15866

Figure Lengend Snippet: (A) The transfection efficiency of the siRNA was analysed in OS cells at different time points after transfection using flow cytometry. (B) Western blot analysis of STAT3 and pSTAT3 expression in OS cells treated with ADSC-conditioned medium. OS cells were pretreated with the STAT3 siRNA (50 nM) for 48 h before treatment with ADSC-conditioned medium for an additional 12 h. STAT3 expression levels were detected by western blotting. OS cells treated with Lipofectamine 2000 alone served as negative controls. (C) OS cell invasion was analysed using transwell assay. OS cells were transfected with the siRNA and treated with ADSC-conditioned medium. OS cells treated with ADSC-conditioned medium alone served as positive controls, and OS cells treated with Lipofectamine 2000 alone served as negative controls. (D) The MMP2 and MMP9 levels in the supernatants were detected using ELISAs. (E) OS cells were treated with the STAT3 siRNA or ADSC-conditioned medium, and MMP2/9 and E-cadherin expression in OS cells was examined by western blotting. OS cells treated with Lipofectamine 2000 alone served as negative controls. The results are expressed as mean±SD. Abbreviation: C Control; L Lipofectamine 2000; S siRNA. * P<0.05, ** P<0.01, *** P<0.001.

Article Snippet: Antibody staining with 100 μL of specific antibodies against human STAT3 (1:100 dilution; R&D Systems) and rabbit monoclonal antibodies against human Ki67, MMP2 and MMP9 (1:100 dilution; ProteinTech groups inc, Chicago, IL) was applied to the cells, which were incubated overnight at 4°C.

Techniques: Transfection, Flow Cytometry, Western Blot, Expressing, Transwell Assay, Control

Journal: Oncotarget

Article Title: Adipose-derived mesenchymal stem cells promote osteosarcoma proliferation and metastasis by activating the STAT3 pathway

doi: 10.18632/oncotarget.15866

Figure Lengend Snippet: (A-B) Cell proliferation was evaluated using the CCK8 assay and EdU analysis. OS cells were transfected with the siRNA and treated with ADSC-conditioned medium. OS cells treated with ADSC-conditioned medium alone served as positive controls, and OS cells treated with Lipofectamine 2000 alone served as negative controls. (C-D) OS cells were transfected with the STAT3 siRNA or treated with ADSC-conditioned medium, and apoptosis rates were determined using flow cytometry. OS cells treated with Lipofectamine 2000 alone served as negative controls. The percentages of Annexin V-positive cells are presented in bar charts. *** P<0.001.

Article Snippet: Antibody staining with 100 μL of specific antibodies against human STAT3 (1:100 dilution; R&D Systems) and rabbit monoclonal antibodies against human Ki67, MMP2 and MMP9 (1:100 dilution; ProteinTech groups inc, Chicago, IL) was applied to the cells, which were incubated overnight at 4°C.

Techniques: CCK-8 Assay, Transfection, Flow Cytometry

Journal: Oncotarget

Article Title: Adipose-derived mesenchymal stem cells promote osteosarcoma proliferation and metastasis by activating the STAT3 pathway

doi: 10.18632/oncotarget.15866

Figure Lengend Snippet: (A) An in vivo imaging system was used to monitor OS xenograft luminescence activity, which represented tumour growth and metastasis. (B) Living Image Software was used to analyse tumour bioluminescence intensity weekly. The quantitation of the normalized image counts is shown. (C) Lungs of the tumour-bearing mice were excised, and the bioluminescence intensity was analysed to determine the level of tumour metastasis in the lungs. (D) Survival curves of the three groups are shown, and the median survival of the OS group was 43 days, which was significantly longer than the survival of the OS + conditioned-medium group (25 days, P<0.01). (E) The immunohistochemical analysis of Ki67, STAT3, MMP2 and MMP9 expression in the orthotopic tumour xenografts is shown. (F) Quantitation of the intensity of Ki67, STAT3 and MMP2/9 staining in the xenografts. Scale bar: 25 μm. * P<0.05, ** P<0.01, *** P<0.001.

Article Snippet: Antibody staining with 100 μL of specific antibodies against human STAT3 (1:100 dilution; R&D Systems) and rabbit monoclonal antibodies against human Ki67, MMP2 and MMP9 (1:100 dilution; ProteinTech groups inc, Chicago, IL) was applied to the cells, which were incubated overnight at 4°C.

Techniques: In Vivo Imaging, Activity Assay, Software, Quantitation Assay, Immunohistochemical staining, Expressing, Staining

Journal: Oncogene

Article Title: Overexpression of a dominant negative form of STAT3 selectively impairs hematopoietic stem cell activity.

doi: 10.1038/sj.onc.1205592

Figure Lengend Snippet: Figure 1 Differentiation-associated differences in STAT3 transcript levels in primitive human hematopoietic cells. CD34+CD387

Article Snippet: For detection of wt or dn STAT3, each membrane was blocked with TTS solution (25 mM Tris, pH 7.4, 0.05% Tween 20, 150 mM NaCl) containing 3% skim milk before being incubated with a polyclonal rabbit anti-mouse STAT3 antibody (K15, Santa Cruz Biotech, Santa Cruz, CA, USA) for 1 h. The membrane was thenwashed three times in TTS solution and incubated with a secondary anti-rat Ig conjugated with horse radish peroxidase (Upstate Biotechnology, Lake Placid, NY, USA), washed again three times and developed using a chemiluminescence developing kit (Amersham Pharmacia Biotech).

Techniques:

Journal: Oncogene

Article Title: Overexpression of a dominant negative form of STAT3 selectively impairs hematopoietic stem cell activity.

doi: 10.1038/sj.onc.1205592

Figure Lengend Snippet: Figure 2 Multiplex-RT – PCR analysis of STAT3 isoform ex- pression in human BM cells. (a) The primer sets used and the sizes of amplified products expected are shown at the top of the figure. (The open bar shows the coding region; the asterisk, the location of the stop codon; and the hatched bar, the untranslated region of the STAT3b transcript). (b) Ethidium bromide-stained gel containing multiplex PCR products for b-actin, STAT3a and STAT3b from RNA obtained from the same human marrow sam- ples analysed in Figure 1. (c) Quantification of results from the multiplex PCR shown in (b). The density of each STAT3 band was normalized against that of b-actin and then the values ob- tained from both were expressed as a proportion of the STAT3a value obtained from the CD34+CD387 cells. d=day

Article Snippet: For detection of wt or dn STAT3, each membrane was blocked with TTS solution (25 mM Tris, pH 7.4, 0.05% Tween 20, 150 mM NaCl) containing 3% skim milk before being incubated with a polyclonal rabbit anti-mouse STAT3 antibody (K15, Santa Cruz Biotech, Santa Cruz, CA, USA) for 1 h. The membrane was thenwashed three times in TTS solution and incubated with a secondary anti-rat Ig conjugated with horse radish peroxidase (Upstate Biotechnology, Lake Placid, NY, USA), washed again three times and developed using a chemiluminescence developing kit (Amersham Pharmacia Biotech).

Techniques: Multiplex Assay, Reverse Transcription Polymerase Chain Reaction, Staining

Journal: Oncogene

Article Title: Overexpression of a dominant negative form of STAT3 selectively impairs hematopoietic stem cell activity.

doi: 10.1038/sj.onc.1205592

Figure Lengend Snippet: Figure 3 Evidence of higher STAT3 protein expression in BM cells from 5-FU-treated mice. Balb/C mice were injected with 150 mg/kg of 5-FU and 4 days later their BM cells along with BM cells from untreated control mice, were lysed in 26 Laemmli buffer for Western analysis using an anti-murine STAT3 anti- body. Each lane shows the results for a lysate obtained from the BM cells of an individual mouse. The identity of the 66 kD band that cross-reacted with the anti STAT3 antibody was not es- tablished. UnTx=untreated

Article Snippet: For detection of wt or dn STAT3, each membrane was blocked with TTS solution (25 mM Tris, pH 7.4, 0.05% Tween 20, 150 mM NaCl) containing 3% skim milk before being incubated with a polyclonal rabbit anti-mouse STAT3 antibody (K15, Santa Cruz Biotech, Santa Cruz, CA, USA) for 1 h. The membrane was thenwashed three times in TTS solution and incubated with a secondary anti-rat Ig conjugated with horse radish peroxidase (Upstate Biotechnology, Lake Placid, NY, USA), washed again three times and developed using a chemiluminescence developing kit (Amersham Pharmacia Biotech).

Techniques: Expressing, Injection, Control, Western Blot

Journal: Oncogene

Article Title: Overexpression of a dominant negative form of STAT3 selectively impairs hematopoietic stem cell activity.

doi: 10.1038/sj.onc.1205592

Figure Lengend Snippet: Figure 4 Retroviral vectors. (a) Schematic illustration of the three STAT3 cDNAs (a, b and a dn form of STAT3). LZ=leu- cine zipper; DNBD=DNA binding domain; SH2=src homology domain 2; TA=transactivating domain. Y indicates the tyrosine residue at position 705 and S indicates the serine residue at posi- tion 727, both of which are key phosphorylation sites. (b) Sche- matic illustration of the mechanism of action of dn STAT3. Upon dimerization of gp130, activated JAK2 phosphorylates a distal tyrosine residue of the intracellular region of gp130. This tyrosine residue then becomes a docking site for the SH2 domain of STAT3, which, in turn, is phosphorylated by JAK2 at Tyr 705. Phosphorylation of Tyr 705 promotes the dimerization of the ‘ac- tivated’ STAT3 molecules that allows them to translocate to the nucleus where they function as promoter-specific activators of transcription. The truncated dn STAT3 can bind to the phos- phorylated tyrosine on activated gp130 but cannot undergo the phosphorylation required for its subsequent activation because it lacks the C-terminal region distal to the SH2 domain (which in- cludes Tyr 705). (c) Schematic diagram of the MSCV-based retro- viral vectors used. These result in expression of GFP only (MIG), GFP plus wild-type (WT) STAT3, or GFP plus dn STAT3. Also shown are the probes used to detect amplified products derived from reverse transcribed RNA isolated from transduced cells. LTR=long terminal repeat (derived from MSCV); SD, SA=spli- cing donor and acceptor sites, respectively, IRES=internal ribo- somal entry site. MIG=MSCV-IRES-GFP, S=Sst1 digestion site

Article Snippet: For detection of wt or dn STAT3, each membrane was blocked with TTS solution (25 mM Tris, pH 7.4, 0.05% Tween 20, 150 mM NaCl) containing 3% skim milk before being incubated with a polyclonal rabbit anti-mouse STAT3 antibody (K15, Santa Cruz Biotech, Santa Cruz, CA, USA) for 1 h. The membrane was thenwashed three times in TTS solution and incubated with a secondary anti-rat Ig conjugated with horse radish peroxidase (Upstate Biotechnology, Lake Placid, NY, USA), washed again three times and developed using a chemiluminescence developing kit (Amersham Pharmacia Biotech).

Techniques: Retroviral, Dominant Negative Mutation, Binding Assay, Residue, Phospho-proteomics, Activation Assay, Expressing, Derivative Assay, Reverse Transcription, Isolation

Journal: Oncogene

Article Title: Overexpression of a dominant negative form of STAT3 selectively impairs hematopoietic stem cell activity.

doi: 10.1038/sj.onc.1205592

Figure Lengend Snippet: Figure 6 Expression of STAT3 proteins in transduced primary hematopoietic cells. Lysates were prepared from aliquots of trans- duced mouse fetal liver cells and analysed for the presence of STAT3 proteins using a polyclonal anti-STAT3 antibody (105 cells per lane). Arrows indicate the expected size of intact gene products. Quantitation of the ratio of STAT3a to STAT3b showed this to be 1.4 in both MIG and wt STAT3-transduced cells

Article Snippet: For detection of wt or dn STAT3, each membrane was blocked with TTS solution (25 mM Tris, pH 7.4, 0.05% Tween 20, 150 mM NaCl) containing 3% skim milk before being incubated with a polyclonal rabbit anti-mouse STAT3 antibody (K15, Santa Cruz Biotech, Santa Cruz, CA, USA) for 1 h. The membrane was thenwashed three times in TTS solution and incubated with a secondary anti-rat Ig conjugated with horse radish peroxidase (Upstate Biotechnology, Lake Placid, NY, USA), washed again three times and developed using a chemiluminescence developing kit (Amersham Pharmacia Biotech).

Techniques: Expressing, Quantitation Assay

Journal: Oncogene

Article Title: Overexpression of a dominant negative form of STAT3 selectively impairs hematopoietic stem cell activity.

doi: 10.1038/sj.onc.1205592

Figure Lengend Snippet: Figure 7 Lack of effect of overexpression of dn STAT3 and WT – STAT3 on the detection of CFU-S and CFCs. Each bar shows the frequency of progenitors measured initially (d0) and immediately post-infection per 104 or 103 TER119+ cell-depleted murine fetal liver cells initially placed in culture, respectively. Va- lues shown are the mean+s.e.m. of data pooled from multiple ex- periments (CFU-S assays, nine mice/group, four experiments; CFC assays, six experiments)

Article Snippet: For detection of wt or dn STAT3, each membrane was blocked with TTS solution (25 mM Tris, pH 7.4, 0.05% Tween 20, 150 mM NaCl) containing 3% skim milk before being incubated with a polyclonal rabbit anti-mouse STAT3 antibody (K15, Santa Cruz Biotech, Santa Cruz, CA, USA) for 1 h. The membrane was thenwashed three times in TTS solution and incubated with a secondary anti-rat Ig conjugated with horse radish peroxidase (Upstate Biotechnology, Lake Placid, NY, USA), washed again three times and developed using a chemiluminescence developing kit (Amersham Pharmacia Biotech).

Techniques: Over Expression, Infection

Journal: Oncogene

Article Title: Overexpression of a dominant negative form of STAT3 selectively impairs hematopoietic stem cell activity.

doi: 10.1038/sj.onc.1205592

Figure Lengend Snippet: Figure 8 In vivo effects of over-expressing wt or dn STAT3 on the long-term lympho-myeloid repopulating ability of murine fetal liver cells. The level of engraftment by transduced (GFP+, left three panels) versus non-transduced (GFP7, right three panels) donor-derived (Ly5.1+) WBCs in individual Ly5.2+ recipients transplanted 5 (top two panels), 14 (middle two panels) and 52 weeks (bottom two panels) previously with equal numbers of Ly5.1+ fetal liver cells is shown (MIG, solid symbols; dn STAT3, open symbols; and WT – STAT3, hatched symbols). Results are pooled from two independent experiments. Each symbol corre- sponds to a single recipient mouse

Article Snippet: For detection of wt or dn STAT3, each membrane was blocked with TTS solution (25 mM Tris, pH 7.4, 0.05% Tween 20, 150 mM NaCl) containing 3% skim milk before being incubated with a polyclonal rabbit anti-mouse STAT3 antibody (K15, Santa Cruz Biotech, Santa Cruz, CA, USA) for 1 h. The membrane was thenwashed three times in TTS solution and incubated with a secondary anti-rat Ig conjugated with horse radish peroxidase (Upstate Biotechnology, Lake Placid, NY, USA), washed again three times and developed using a chemiluminescence developing kit (Amersham Pharmacia Biotech).

Techniques: In Vivo, Expressing, Derivative Assay

Journal: Oncogene

Article Title: Overexpression of a dominant negative form of STAT3 selectively impairs hematopoietic stem cell activity.

doi: 10.1038/sj.onc.1205592

Figure Lengend Snippet: Figure 9 Expression of transduced genes in the regenerated WBCs of mice transplanted with fetal liver cells infected with MIG, wt STAT3 and dn STAT3 vectors. WBCs were obtained from individual recipient mice 5 weeks after they had been trans- planted and analysed by RT – PCR for the presence of the expected size of vector-derived transcript (indicated in Figure 4(c)). The re- sultant ethidium bromide-stained products are shown in the top panel and the GFP-probed Southern blots of the same gels are shown in the middle. Results for b-actin are shown in the bottom panel. Each lane shows the results for a single mouse whose per cent GFP+ WBCs is indicated in brackets. Lanes labeled MIG/ DNA, dnS/DNA and WT/DNA were loaded with PCR products from plasmids containing the MIG, dn STAT3 and wt STAT3 vec- tors as positive controls. Also included as positive controls were RT – PCR products from RNA extracts of the corresponding GPE-86 producer cells (MIG, GPE-dnS and GPE-WT)

Article Snippet: For detection of wt or dn STAT3, each membrane was blocked with TTS solution (25 mM Tris, pH 7.4, 0.05% Tween 20, 150 mM NaCl) containing 3% skim milk before being incubated with a polyclonal rabbit anti-mouse STAT3 antibody (K15, Santa Cruz Biotech, Santa Cruz, CA, USA) for 1 h. The membrane was thenwashed three times in TTS solution and incubated with a secondary anti-rat Ig conjugated with horse radish peroxidase (Upstate Biotechnology, Lake Placid, NY, USA), washed again three times and developed using a chemiluminescence developing kit (Amersham Pharmacia Biotech).

Techniques: Expressing, Infection, Reverse Transcription Polymerase Chain Reaction, Plasmid Preparation, Derivative Assay, Staining, Labeling

Journal: Cancer biology & therapy

Article Title: Lack of AKT activation in lung cancer cells with EGFR mutation is a novel marker of cetuximab sensitivity.

doi: 10.4161/cbt.19238

Figure Lengend Snippet: Figure 2. Phosphorylation of EGFR, ERK, AKT and STAT3, and expression and mutation status of PTEN in EGFR mutant and wild-type NSCLC cell lines. (A–D) Cells were incubated with/without serum for 24 h, and immunoblotting was done with antibod- ies targeting phosphorylated (p) or total EGFR, ERK and AKT. Densitometric analysis was performed with TotaLab software (Nonlinear Dynamics), and the rela- tive pEGFR/EGFR, pERK/ERK, pAKT/AKT, pSTAT3/STAT3 ratios are indicated as bar graphs under each blot. Experiments were repeated three times and represen- tative results are shown. (E) PTEN protein status in each cell line was determined by immunoblotting in the presence or absence of serum. The β-actin was used as loading control. Experiments were repeated three times and representative results are shown. The muta- tion status of PTEN in each cell line is also indicated. WT, wild type.

Article Snippet: The following primary antibodies were used: pEGFR (Tyr 1068), pAKT (Ser 473), AKT, pSTAT3 (Tyr 705), STAT3, PTEN (Cell Signaling, MA), EGFR (1005) (Santa Cruz), β-ACTIN (Sigma), pERK and ERK (Promega).

Techniques: Phospho-proteomics, Expressing, Mutagenesis, Incubation, Western Blot, Software, Control

Journal: Pharmaceutics

Article Title: Exosomes Derived from BMSCs Treated with CeONPs Ameliorate Radiation-Induced Jaw Bone Injury via miR-21-5p/STAT3 Axis-Mediated Osteogenesis and ROS Scavenging

doi: 10.3390/pharmaceutics18020216

Figure Lengend Snippet: miR-21-5p derived from BMSC-Ce-exos regulates STAT3 (wt: wild-type; mut: mutant; NC: negative control). ( A , B ) Heatmap and volcano plot of miRNA sequencing analysis, with blue indicating downregulated miRNAs and red indicating upregulated ones; ( C ) qRT-PCR analysis of the top five expressed miRNAs (miR-140-3p, miR-21-5p, miR-143-3p, miR-27b-3p, and miR-34c-5p) in BMSC-Ce-exos; ( D ) PPI network analysis of intersecting target genes; ( E ) String analysis of the PPI network interaction results; ( F ) Binding sequence of miR-21-5p in the 3′-UTR of STAT3; ( G ) Luciferase readout from BMSCs co-transfected with wt or mut STAT3 3′-UTR and control mimics or miR-21-5p mimics; ( H ) Protein expression of STAT3 in BMSCs transfected with miR-21-5p mimics, miR-21-5p inhibitor and their NCs; ( I ) mRNA expression levels of STAT3 transfected with miR-21-5p mimics, miR-21-5p inhibitor and their NCs in BMSCs. (ns) p > 0.05; (*) p < 0.05; (**) p < 0.01, and (***) p < 0.001.

Article Snippet: Subsequently, 20 μg of proteins were separated via 4–20% SDS-PAGE at 160 V for 40 min and transferred onto polyvinylidene fluoride (PVDF) membranes at 400 mA for 30 min. Membranes were blocked with 5% non-fat milk and incubated overnight at 4 °C with primary antibodies against GAPDH (1:5000, HRP-60004, Proteintech, Chicago, IL, USA), ALP (1:500, ab65834, Abcam, Cambridge, UK), Runx2 (1:1000, GTX00792, GeneTex, Irvine, CA, USA), ColA1 (1:1000, GTX112731, GeneTex), OCN (1:1000, DF12303, Affinity, Cincinnati, OH, USA), TSG101 (1:1000, ab125011, Abcam), CD63 (1:1000, 67605-1-lg, Proteintech), Flotillin-2 (1:1000, 66881-1-lg, Proteintech), STAT3 (1:1000, 10253-2-ap, Proteintech), Nrf2 (1:1000, GB113808 , Servicebio, Wuhan, China), Calnexin (1:1000, ab22595, Abcam), and β-Actin (1:1000, GTX109639, GeneTex).

Techniques: Derivative Assay, Mutagenesis, Negative Control, Sequencing, Quantitative RT-PCR, Binding Assay, Luciferase, Transfection, Control, Expressing

Journal: Pharmaceutics

Article Title: Exosomes Derived from BMSCs Treated with CeONPs Ameliorate Radiation-Induced Jaw Bone Injury via miR-21-5p/STAT3 Axis-Mediated Osteogenesis and ROS Scavenging

doi: 10.3390/pharmaceutics18020216

Figure Lengend Snippet: Exosomal miR-21-5p promotes osteogenesis and reduces the ROS levels by targeting STAT3. ( A ) The red staining indicative of mineralization is most obvious when BMSCs are cotransfected with miR-21-5p mimics and pcDNA-NC, while matrix mineralization is significantly reduced when BMSCs are cotransfected with miR-NC and pcDNA-STAT3; ( B ) Quantitative analysis shows that overexpression of STAT3 prevents the enhancement of osteoblastic differentiation in BMSCs induced by miR-21-5p mimics; ( C ) DHE immunofluorescence shows the ROS levels among different groups; ( D ) Quantitative analysis indicates that overexpression of STAT3 prevents the alleviation of ROS accumulation in BMSCs by miR-21-5p mimics; ( E ) SOD assay shows superoxide dismutase levels among different groups; ( F ) MDA assay shows lipid peroxide levels among different groups; ( G ) Western blotting assays shows that overexpression of STAT3 prevents the upregulation of Runx2, Col1a1, ALP, and OCN protein expression induced by miR-21-5p mimics; ( H ) Western blotting assays shows that overexpression of STAT3 prevents the upregulation of Nrf2 protein expression induced by miR-21-5p mimics. (**) p < 0.01; (****) p < 0.0001.

Article Snippet: Subsequently, 20 μg of proteins were separated via 4–20% SDS-PAGE at 160 V for 40 min and transferred onto polyvinylidene fluoride (PVDF) membranes at 400 mA for 30 min. Membranes were blocked with 5% non-fat milk and incubated overnight at 4 °C with primary antibodies against GAPDH (1:5000, HRP-60004, Proteintech, Chicago, IL, USA), ALP (1:500, ab65834, Abcam, Cambridge, UK), Runx2 (1:1000, GTX00792, GeneTex, Irvine, CA, USA), ColA1 (1:1000, GTX112731, GeneTex), OCN (1:1000, DF12303, Affinity, Cincinnati, OH, USA), TSG101 (1:1000, ab125011, Abcam), CD63 (1:1000, 67605-1-lg, Proteintech), Flotillin-2 (1:1000, 66881-1-lg, Proteintech), STAT3 (1:1000, 10253-2-ap, Proteintech), Nrf2 (1:1000, GB113808 , Servicebio, Wuhan, China), Calnexin (1:1000, ab22595, Abcam), and β-Actin (1:1000, GTX109639, GeneTex).

Techniques: Staining, Over Expression, Immunofluorescence, Multiple Displacement Amplification, Western Blot, Expressing

Journal: Virology Journal

Article Title: Influenza a virus regulates interferon signaling and its associated genes; MxA and STAT3 by cellular miR-141 to ensure viral replication

doi: 10.1186/s12985-023-02146-4

Figure Lengend Snippet: Oligonucleotides sequences used for quantifying miR-141 and mRNA of indicated genes

Article Snippet: For staining MxA and STAT3, the primary antibodies; rabbit polyclonal anti-MxA (Novus Biologicals, NBP132905) and mouse monoclonal anti-STAT3 (Abcam, ab119352) were used.

Techniques:

Journal: Virology Journal

Article Title: Influenza a virus regulates interferon signaling and its associated genes; MxA and STAT3 by cellular miR-141 to ensure viral replication

doi: 10.1186/s12985-023-02146-4

Figure Lengend Snippet: The correlation between miR-141 level and the expression profile of MxA and STAT3 in infected A549 cells. (A) Quantification of steady-state miR-141 in infected A549 cells with MOI of 0.5 and transfected with either pre-miR-141 or miR-141 inhibitor compared with noninfected cells (control) using qRT-PCR. (B) Relative gene expression of MxA and STAT3 in infected A549 cells transfected with either specific inhibitor against miR-141 or pre- miR-141 compared with control-transfected cells using qRT-PCR. Error bars indicate the STD of three independent experiments. Student two-tailed t -test used for statistical analysis, (*) indicates P-values ≤ 0.05, and (**) indicates P ≤ 0.01. (C) Flow cytometric assay quantifies the kinetic proteins expression profile of MxA (in blue dots) and STAT3 (in red dots) in infected and transfected A549 cells compared with control cells. (D) Western blot analysis reveals the protein expression level of MxA and STAT3 in infected and transfected cells compared to control cells, β-actin expression profile severed as an internal control

Article Snippet: For staining MxA and STAT3, the primary antibodies; rabbit polyclonal anti-MxA (Novus Biologicals, NBP132905) and mouse monoclonal anti-STAT3 (Abcam, ab119352) were used.

Techniques: Expressing, Infection, Transfection, Control, Quantitative RT-PCR, Gene Expression, Two Tailed Test, Flow Cytometry, Western Blot

Journal: Virology Journal

Article Title: Influenza a virus regulates interferon signaling and its associated genes; MxA and STAT3 by cellular miR-141 to ensure viral replication

doi: 10.1186/s12985-023-02146-4

Figure Lengend Snippet: Quantification analysis of miR-141, MxA, and STAT3 in transfected and infected A549 cells

Article Snippet: For staining MxA and STAT3, the primary antibodies; rabbit polyclonal anti-MxA (Novus Biologicals, NBP132905) and mouse monoclonal anti-STAT3 (Abcam, ab119352) were used.

Techniques: Transfection, Infection, Expressing, Control