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Proteintech rabbit polyclonal anti ifitm3
Rabbit Polyclonal Anti Ifitm3, supplied by Proteintech, used in various techniques. Bioz Stars score: 95/100, based on 172 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/rabbit+polyclonal+anti+ifitm3/IFITM3+Antibody/pmc13016079-3-0-4
Average 95 stars, based on 172 article reviews

rabbit polyclonal anti ifitm3 - by Bioz Stars, 2026-10

95/100 stars

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Article Title: Transmembrane domain of IFITM3 is responsible for its interaction with influenza virus HA 2 subunit
Article Snippet: The protein bands were detected by Western blotting HPR substrate (Millipore, Germany) using the FluorChem imagers (Protein-simple, USA). .. The primary antibodies used in this study include Rabbit polyclonal anti-IFITM3 (Proteintech, Cat# 11714-1-AP, USA), FLAG-tag Polyclonal Antibody (MultiSciences, Cat# ab30107, China), β-Actin Mouse mAb (MultiSciences, Cat# ab008, China), Anti-HA (H5N1) (Avian) (Immune Technology, Cat# IT-003-006, USA), Anti-HA (H7N9) (A/Shanghai/1/2013) (Immune Technology, Cat# IT-003-0073M1, USA), Anti-EGFP antibody (F56-6A1.2.3) (Abcam, Cat# ab184601, UK), Anti-mCherry antibody (Abcam, Cat# ab167453, UK), GAPDH (14C10) Rabbit mAb (Cell Signaling Technology, Cat# 2118, USA), and Myc-tag Polyclonal Antibody (MultiSciences, Cat# ab30104, China). .. The secondary antibodies used in this study include Goat Anti-Rabbit IgG (HRP) (Beyotime, Cat# A0208, China) and Goat Anti-Rabbit IgG (HRP) (Beyotime, Cat# A0216, China).

Article Title: Transmembrane domain of IFITM3 is responsible for its interaction with influenza virus HA 2 subunit.
Article Snippet: The protein bands were detected by Western blotting HPR substrate (Millipore, Germany) using the FluorChem imagers (Protein-simple, USA). .. The primary antibodies used in this study include Rabbit polyclonal anti-IFITM3 (Proteintech, Cat# 11714-1-AP, USA), FLAG-tag Polyclonal Antibody (MultiSciences, Cat# ab30107, China), β-Actin Mouse mAb (MultiSciences, Cat# ab008, China), Anti-HA (H5N1) (Avian) (Immune Technology, Cat# IT-003-006, USA), Anti-HA (H7N9) (A/Shanghai/1/ 2013) (Immune Technology, Cat# IT-003-0073M1, USA), Anti-EGFP antibody (F56-6A1.2.3) (Abcam, Cat# ab184601, UK), Anti-mCherry antibody (Abcam, Cat# ab167453, UK), GAPDH (14C10) Rabbit mAb (Cell Signaling Technology, Cat# 2118, USA), and Myc-tag Polyclonal Antibody (MultiSciences, Cat# ab30104, China). .. The secondary antibodies used in this study include Goat Anti-Rabbit IgG (HRP) (Beyotime, Cat# A0208, China) and Goat Anti-Rabbit IgG (HRP) (Beyotime, Cat# A0216, China).

other:

Article Title: MYCT1–IFITM2/3 interaction links endothelial endolysosomal trafficking to white adipose tissue expansion
Article Snippet: Rabbit polyclonal anti-IFITM3 , Proteintech , Cat# 11714-1-AP, RRID:AB_2295684.

Image Search Results

Overexpression of IFITM3 in LPS-stimulated BV2 cells enhances inflammatory responses. ( A ) Venn diagram of the GSE73878 and GSE18740 datasets. ( B ) RT-qPCR analysis demonstrated that IFITM3 was significantly upregulated in the cellular inflammation model. ( C ) The knockdown efficiency of si-IFITM3-1 to -3 was measured by RT-qPCR. ( D ) RT-qPCR analyses and ( E ) western blot were used to detect the effect of IFITM3 overexpression. ( F – H ) Suppression of IFITM3 expression reduced the production of IL-1β, IL-6, and TNF-α in LPS-treated BV2 cells. ( I – K ) The production of IL-1β, IL-6, and TNF-α was enhanced following transfection of IFITM3 into BV2 cells treated with LPS. ( L – N ) The levels of IL-1β, IL-6, and TNF-α were measured using ELISA in LPS-treated BV2 cells transfected with si-IFITM3-2, si-IFITM3-3, IFITM3, or their corresponding negative controls.

Journal: Scientific Reports

Article Title: IFITM3-mediated neuroinflammation in epilepsy regulated by the let-7g-5p/STAT2 axis

doi: 10.1038/s41598-026-44357-z

Figure Lengend Snippet: Overexpression of IFITM3 in LPS-stimulated BV2 cells enhances inflammatory responses. ( A ) Venn diagram of the GSE73878 and GSE18740 datasets. ( B ) RT-qPCR analysis demonstrated that IFITM3 was significantly upregulated in the cellular inflammation model. ( C ) The knockdown efficiency of si-IFITM3-1 to -3 was measured by RT-qPCR. ( D ) RT-qPCR analyses and ( E ) western blot were used to detect the effect of IFITM3 overexpression. ( F – H ) Suppression of IFITM3 expression reduced the production of IL-1β, IL-6, and TNF-α in LPS-treated BV2 cells. ( I – K ) The production of IL-1β, IL-6, and TNF-α was enhanced following transfection of IFITM3 into BV2 cells treated with LPS. ( L – N ) The levels of IL-1β, IL-6, and TNF-α were measured using ELISA in LPS-treated BV2 cells transfected with si-IFITM3-2, si-IFITM3-3, IFITM3, or their corresponding negative controls.

Article Snippet: The primary antibodies targeted STAT2 (rabbit, 1:100; Proteintech) and IFITM3 (mouse, 1:100; Zhongshan Golden Bridge, Beijing, China).

Techniques: Over Expression, Quantitative RT-PCR, Knockdown, Western Blot, Expressing, Transfection, Enzyme-linked Immunosorbent Assay

IFITM3 promotes apoptosis and pyroptosis in LPS-treated BV2 cells. ( A ) The rate of BV2 cells undergoing pyroptosis was quantified by flow cytometry. ( B , C ) The relative expression levels of ASC and IL-18 were determined by RT-qPCR.

Journal: Scientific Reports

Article Title: IFITM3-mediated neuroinflammation in epilepsy regulated by the let-7g-5p/STAT2 axis

doi: 10.1038/s41598-026-44357-z

Figure Lengend Snippet: IFITM3 promotes apoptosis and pyroptosis in LPS-treated BV2 cells. ( A ) The rate of BV2 cells undergoing pyroptosis was quantified by flow cytometry. ( B , C ) The relative expression levels of ASC and IL-18 were determined by RT-qPCR.

Article Snippet: The primary antibodies targeted STAT2 (rabbit, 1:100; Proteintech) and IFITM3 (mouse, 1:100; Zhongshan Golden Bridge, Beijing, China).

Techniques: Flow Cytometry, Expressing, Quantitative RT-PCR

IFITM3 knockdown alleviates seizure severity and neuroinflammatory responses in PTZ-kindled mice. ( A ) Western blot analysis was conducted to assess the expression of IFITM3, verifying the knockdown efficiency of LV-sh-IFITM3 in BV2 cells treated with LPS ( n = 3). ( B ) Severity of seizures was scored according to the Racine scale for the vehicle + LV-sh-NC, PTZ + LV-sh-NC, and PTZ + LV-sh-IFITM3 groups. No seizures were observed in the vehicle + LV-sh-NC group, yielding a score of 0 ( n = 16). ( C ) Representative EEG traces recorded in vivo from each experimental group ( n = 3). ( D – I ) mRNA levels of IFITM3, IL-1β, IL-6, TNF-α, IL-18, and ASC in mouse cortical tissue were determined by RT-qPCR ( n = 16). ( J , K ) Protein levels of IFITM3, cleaved caspase-3, caspase-3, cleaved caspase9, caspase-9, BAX, and BCL2 were evaluated by western blotting in the cortex lysates ( n = 3). Statistical analysis was conducted using two-way ANOVA followed by Bonferroni multiple comparison tests. Data are presented as mean ± standard deviation.

Journal: Scientific Reports

Article Title: IFITM3-mediated neuroinflammation in epilepsy regulated by the let-7g-5p/STAT2 axis

doi: 10.1038/s41598-026-44357-z

Figure Lengend Snippet: IFITM3 knockdown alleviates seizure severity and neuroinflammatory responses in PTZ-kindled mice. ( A ) Western blot analysis was conducted to assess the expression of IFITM3, verifying the knockdown efficiency of LV-sh-IFITM3 in BV2 cells treated with LPS ( n = 3). ( B ) Severity of seizures was scored according to the Racine scale for the vehicle + LV-sh-NC, PTZ + LV-sh-NC, and PTZ + LV-sh-IFITM3 groups. No seizures were observed in the vehicle + LV-sh-NC group, yielding a score of 0 ( n = 16). ( C ) Representative EEG traces recorded in vivo from each experimental group ( n = 3). ( D – I ) mRNA levels of IFITM3, IL-1β, IL-6, TNF-α, IL-18, and ASC in mouse cortical tissue were determined by RT-qPCR ( n = 16). ( J , K ) Protein levels of IFITM3, cleaved caspase-3, caspase-3, cleaved caspase9, caspase-9, BAX, and BCL2 were evaluated by western blotting in the cortex lysates ( n = 3). Statistical analysis was conducted using two-way ANOVA followed by Bonferroni multiple comparison tests. Data are presented as mean ± standard deviation.

Article Snippet: The primary antibodies targeted STAT2 (rabbit, 1:100; Proteintech) and IFITM3 (mouse, 1:100; Zhongshan Golden Bridge, Beijing, China).

Techniques: Knockdown, Western Blot, Expressing, In Vivo, Quantitative RT-PCR, Comparison, Standard Deviation

STAT2 as a potential transcription factor for IFITM3. ( A ) The expression levels of transcription factors IRF9, STAT2, MAFF, TFAP2C, and PITX2 in BV2 cells were measured, with Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) serving as the internal control. The overexpression efficiency of ( B ) IRF9 or ( C ) STAT2 plasmids was measured. ( D ) The inhibition efficiency of four si-IRF9 constructs was assessed in BV2 cells using RT-qPCR. ( E ) The inhibition efficiency of four si-IFITM3 constructs was similarly evaluated in BV2 cells. IFITM3 expression was evaluated by RT-qPCR in BV2 cells transfected with ( F ) IRF9 or ( G ) STAT2. RT-qPCR was performed to quantify IFITM3 expression in BV2 cells transfected with ( H ) si-IRF9-1 or ( I ) si-STAT2-3. ( J ) HEK293T cells were co-transfected with the IFITM3 promoter and negative control, IRF9, or STAT2 plasmids. After 48 h, luciferase activity and corresponding protein expression were detected. ( K ) BV2 cells were co-transfected with STAT2 or negative control together with either the IFITM3 promoter or IFITM3-promoter-mut. The relative luciferase activity was measured. ( L ) IFITM3 protein levels were examined by western blot in BV2 cells transfected with si-NC, si-STAT2-3, si-STAT2-4, oe-NC or STAT2. ( M ) Representative immunofluorescence images (magnification ×400) demonstrate the expression of IFITM3. Blue: nuclear staining (DAPI); red: IFITM3 staining. Scale bar: 100 μm. STAT2 expression in the cortex ( N ) and hippocampus ( O ) of epileptic mice was assessed by RT-qPCR ( n = 16).

Journal: Scientific Reports

Article Title: IFITM3-mediated neuroinflammation in epilepsy regulated by the let-7g-5p/STAT2 axis

doi: 10.1038/s41598-026-44357-z

Figure Lengend Snippet: STAT2 as a potential transcription factor for IFITM3. ( A ) The expression levels of transcription factors IRF9, STAT2, MAFF, TFAP2C, and PITX2 in BV2 cells were measured, with Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) serving as the internal control. The overexpression efficiency of ( B ) IRF9 or ( C ) STAT2 plasmids was measured. ( D ) The inhibition efficiency of four si-IRF9 constructs was assessed in BV2 cells using RT-qPCR. ( E ) The inhibition efficiency of four si-IFITM3 constructs was similarly evaluated in BV2 cells. IFITM3 expression was evaluated by RT-qPCR in BV2 cells transfected with ( F ) IRF9 or ( G ) STAT2. RT-qPCR was performed to quantify IFITM3 expression in BV2 cells transfected with ( H ) si-IRF9-1 or ( I ) si-STAT2-3. ( J ) HEK293T cells were co-transfected with the IFITM3 promoter and negative control, IRF9, or STAT2 plasmids. After 48 h, luciferase activity and corresponding protein expression were detected. ( K ) BV2 cells were co-transfected with STAT2 or negative control together with either the IFITM3 promoter or IFITM3-promoter-mut. The relative luciferase activity was measured. ( L ) IFITM3 protein levels were examined by western blot in BV2 cells transfected with si-NC, si-STAT2-3, si-STAT2-4, oe-NC or STAT2. ( M ) Representative immunofluorescence images (magnification ×400) demonstrate the expression of IFITM3. Blue: nuclear staining (DAPI); red: IFITM3 staining. Scale bar: 100 μm. STAT2 expression in the cortex ( N ) and hippocampus ( O ) of epileptic mice was assessed by RT-qPCR ( n = 16).

Article Snippet: The primary antibodies targeted STAT2 (rabbit, 1:100; Proteintech) and IFITM3 (mouse, 1:100; Zhongshan Golden Bridge, Beijing, China).

Techniques: Expressing, Control, Over Expression, Inhibition, Construct, Quantitative RT-PCR, Transfection, Negative Control, Luciferase, Activity Assay, Western Blot, Immunofluorescence, Staining

STAT2 promotes inflammation, apoptosis, and pyroptosis through modulation of IFITM3. ( A – C ) Levels of IL-1β, IL-6, and TNF-α were quantified in BV2 cells transfected with STAT2, si-IFITM3-3, or the corresponding negative controls. The rescue experiments involved co-transfection of STAT2 with si-IFITM3-3. ( D – F ) The expression levels of IL-1β, IL-6, and TNF-α in BV2 cells transfected with si-STAT2-3, IFITM3, or the respective negative controls. The rescue experiments involved co-transfection of si-STAT2-3 with IFITM3. ( G ) The apoptosis rate of BV2 cells transfected with si-STAT2-3, IFITM3, or their negative controls. ( H ) Flow cytometry was performed to assess apoptosis rate in each treatment group. The expression levels of ( I , K ) ASC and ( J , L ) IL-18 in BV2 cells treated with different plasmids were detected by RT-qPCR analysis.

Journal: Scientific Reports

Article Title: IFITM3-mediated neuroinflammation in epilepsy regulated by the let-7g-5p/STAT2 axis

doi: 10.1038/s41598-026-44357-z

Figure Lengend Snippet: STAT2 promotes inflammation, apoptosis, and pyroptosis through modulation of IFITM3. ( A – C ) Levels of IL-1β, IL-6, and TNF-α were quantified in BV2 cells transfected with STAT2, si-IFITM3-3, or the corresponding negative controls. The rescue experiments involved co-transfection of STAT2 with si-IFITM3-3. ( D – F ) The expression levels of IL-1β, IL-6, and TNF-α in BV2 cells transfected with si-STAT2-3, IFITM3, or the respective negative controls. The rescue experiments involved co-transfection of si-STAT2-3 with IFITM3. ( G ) The apoptosis rate of BV2 cells transfected with si-STAT2-3, IFITM3, or their negative controls. ( H ) Flow cytometry was performed to assess apoptosis rate in each treatment group. The expression levels of ( I , K ) ASC and ( J , L ) IL-18 in BV2 cells treated with different plasmids were detected by RT-qPCR analysis.

Article Snippet: The primary antibodies targeted STAT2 (rabbit, 1:100; Proteintech) and IFITM3 (mouse, 1:100; Zhongshan Golden Bridge, Beijing, China).

Techniques: Transfection, Cotransfection, Expressing, Flow Cytometry, Quantitative RT-PCR

Analyzing the knockdown of IFITM3 efficiency in LV-shIFITM3 KG-1a cells. (A) The efficiency of lentivirus infection of the KG-1a cells was observed under a conventional fluorescent microscope. (B) The IFITM3 mRNA expression in the KG-1a cells was detected by qRT-PCR. (C) The expression of the IFITM3 protein in the KG-1a cells was detected by Western blot. (D) The results from the semi-quantitative analysis of protein expression levels. ****P<0.00001. ns, no statistical significance.

Journal: Frontiers in Oncology

Article Title: IONPs combined with cytarabine downregulated IFITM3 expression to inhibit acute myeloid leukemia

doi: 10.3389/fonc.2025.1515956

Figure Lengend Snippet: Analyzing the knockdown of IFITM3 efficiency in LV-shIFITM3 KG-1a cells. (A) The efficiency of lentivirus infection of the KG-1a cells was observed under a conventional fluorescent microscope. (B) The IFITM3 mRNA expression in the KG-1a cells was detected by qRT-PCR. (C) The expression of the IFITM3 protein in the KG-1a cells was detected by Western blot. (D) The results from the semi-quantitative analysis of protein expression levels. ****P<0.00001. ns, no statistical significance.

Article Snippet: The membrane was then incubated with the rabbit anti-human GAPDH monoclonal antibody and the rabbit anti-human IFITM3 polyclonal antibody (Wuhan Sanying Biotechnology Co. LTD), respectively, for overnight at 4°C.

Techniques: Knockdown, Infection, Microscopy, Expressing, Quantitative RT-PCR, Western Blot

Down-regulation of IFITM3 changed the biological properties of the KG-1a cells. (A) The effect of down-regulated IFITM3 on the proliferation ability of KG-1a cells as detected by CCK8 assay. (B) The colony formations of various KG-1a cells detected by soft agar cloning assay (50×); (C) Comparisons of the colony formations; (D) The FCM analysis results of the KG-1a cell cycle changes; (E) Comparisons of the cell cycles; (F) The effects of Ara-C (0.4μM) on the apoptosis of various KG-1a cells analyzed by FCM (24h). (G) The FCM results; (H) The mRNA expression level of caspase 3 in various KG-1a cells as detected by qRT-PCR. *P < 0.05, ***P < 0.001. ns, no statistical significance.

Journal: Frontiers in Oncology

Article Title: IONPs combined with cytarabine downregulated IFITM3 expression to inhibit acute myeloid leukemia

doi: 10.3389/fonc.2025.1515956

Figure Lengend Snippet: Down-regulation of IFITM3 changed the biological properties of the KG-1a cells. (A) The effect of down-regulated IFITM3 on the proliferation ability of KG-1a cells as detected by CCK8 assay. (B) The colony formations of various KG-1a cells detected by soft agar cloning assay (50×); (C) Comparisons of the colony formations; (D) The FCM analysis results of the KG-1a cell cycle changes; (E) Comparisons of the cell cycles; (F) The effects of Ara-C (0.4μM) on the apoptosis of various KG-1a cells analyzed by FCM (24h). (G) The FCM results; (H) The mRNA expression level of caspase 3 in various KG-1a cells as detected by qRT-PCR. *P < 0.05, ***P < 0.001. ns, no statistical significance.

Techniques: CCK-8 Assay, Cloning, Expressing, Quantitative RT-PCR

The IFITM3 expression and the proliferation of the KG-1a cells. (A) The IFITM3 expression as detected by Western blot after the KG-1a cells were treated with PBS, PBNPs and IONPs, respectively, for 72h; (B) The semi-quantitative analysis of the Western blot results; (C) The IFITM3 expression in the KG-1a cells as detected by Western blot after the cells were treated with PBS, PBNPs and IONPs, respectively, for 72h; (D) The proliferation of the KG-1a cells as detected by the CCK8 assay in the KG-1a cells that were treated with different drugs for 96h; (E) The qRT-PCR-detected expression level of c-myc in the KG-1a cells treated with different drugs for 24h. *P < 0.05, **P < 0.01, ***P < 0.001. ns, no statistical significance.

Journal: Frontiers in Oncology

Article Title: IONPs combined with cytarabine downregulated IFITM3 expression to inhibit acute myeloid leukemia

doi: 10.3389/fonc.2025.1515956

Figure Lengend Snippet: The IFITM3 expression and the proliferation of the KG-1a cells. (A) The IFITM3 expression as detected by Western blot after the KG-1a cells were treated with PBS, PBNPs and IONPs, respectively, for 72h; (B) The semi-quantitative analysis of the Western blot results; (C) The IFITM3 expression in the KG-1a cells as detected by Western blot after the cells were treated with PBS, PBNPs and IONPs, respectively, for 72h; (D) The proliferation of the KG-1a cells as detected by the CCK8 assay in the KG-1a cells that were treated with different drugs for 96h; (E) The qRT-PCR-detected expression level of c-myc in the KG-1a cells treated with different drugs for 24h. *P < 0.05, **P < 0.01, ***P < 0.001. ns, no statistical significance.

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

Impact of IFITM3 knockdown on disease progression in AML model mice. (A) Leukemia cells in peripheral blood and bone marrow; (B) With Reichsen-Giemsa staining; (C) Leukemia cells of bone marrow (left: under a light microscope; right: under a fluorescence microscope) in the AML-bearing mice injected with the Scrambled or the Lv-shIFITM3-KG1a cells; (D) The expression levels of CD33, CD123 and CD11b detected by FCM. (E) Statistical results of the CD33, CD123 and CD11b expression levels. The experiment was repeated twice. *P < 0.05, ***P < 0.001.

Journal: Frontiers in Oncology

Article Title: IONPs combined with cytarabine downregulated IFITM3 expression to inhibit acute myeloid leukemia

doi: 10.3389/fonc.2025.1515956

Figure Lengend Snippet: Impact of IFITM3 knockdown on disease progression in AML model mice. (A) Leukemia cells in peripheral blood and bone marrow; (B) With Reichsen-Giemsa staining; (C) Leukemia cells of bone marrow (left: under a light microscope; right: under a fluorescence microscope) in the AML-bearing mice injected with the Scrambled or the Lv-shIFITM3-KG1a cells; (D) The expression levels of CD33, CD123 and CD11b detected by FCM. (E) Statistical results of the CD33, CD123 and CD11b expression levels. The experiment was repeated twice. *P < 0.05, ***P < 0.001.

Techniques: Knockdown, Biomarker Discovery, Staining, Light Microscopy, Fluorescence, Microscopy, Injection, Expressing

The over-expression of IFITM3 reduces PRRSV replication. ( a ) Western blotting analysis showing the expression of exogenous IFITM3 in cells transfected with a plasmid containing IFITM3 (I), but not in cells transfected with vector control (V). ( b ) Virus titers in the supernatants of cells transfected with vector control or IFITM3 containing plasmid at 24 h after virus infection. An average of a 5.4-fold decrease was observed in IFITM3-transfected cells compared to vector controls. ( c ) CCK-8 cytotoxicity assay was used to compare the difference in cell viability between QCXIP vector control and IFITM3-HA over-expressing MARC-145 cells. A p -value < 0.05 was considered statistically significant.

Journal: Microorganisms

Article Title: Interferon-Induced Transmembrane Protein 3 (IFITM3) Restricts PRRSV Replication via Post-Entry Mechanisms

doi: 10.3390/microorganisms13081737

Figure Lengend Snippet: The over-expression of IFITM3 reduces PRRSV replication. ( a ) Western blotting analysis showing the expression of exogenous IFITM3 in cells transfected with a plasmid containing IFITM3 (I), but not in cells transfected with vector control (V). ( b ) Virus titers in the supernatants of cells transfected with vector control or IFITM3 containing plasmid at 24 h after virus infection. An average of a 5.4-fold decrease was observed in IFITM3-transfected cells compared to vector controls. ( c ) CCK-8 cytotoxicity assay was used to compare the difference in cell viability between QCXIP vector control and IFITM3-HA over-expressing MARC-145 cells. A p -value < 0.05 was considered statistically significant.

Article Snippet: The following primary antibodies were used: anti-HA mouse monoclonal 1o Ab (6E2) (Cell Signaling Technology, catalog# 2367) at 1:1000 dilution, anti-HA mouse monoclonal antibody (HA7) (Sigma Aldrich, catalog# H3663, Saint Louis, MO, USA) at 1:5000 dilution, anti-PRRSV SR-30 1o monoclonal Ab (provided by Dr. Eric Nelson) at 1:300 dilution, anti-IFITM3 rabbit polyclonal antibody (Proteintech, catalog# 11714-1-AP, Rosemont, IL, USA) at 1:1000 dilution, anti-p-Akt (S743, Cell Signaling Technology, catalog# 9271S) at 1:1000 dilution, anti-LC3B antibody (Sigma Aldrich, catalog# L7542) at 1:2000 dilution, and mouse monoclonal anti-beta actin Ab (Sigma Aldrich, catalog# A2228) at 1:5000 dilution.

Techniques: Over Expression, Western Blot, Expressing, Transfection, Plasmid Preparation, Control, Virus, Infection, CCK-8 Assay, Cytotoxicity Assay

The silencing of IFITM3 slightly enhances PRRSV replication. ( a ) qRT-PCR showing the silencing of IFITM3. The averages and standard deviations of three replicates are shown. ( b ) qRT-PCR showing the viral RNA copies in control silencing and IFITM3 silencing RNA-transfected cells. The averages and standard deviations of three replicates are shown. ( c ) A slightly enhanced TCID 50 virus titer (1.22-fold increase) in IFITM3 silencing RNA-transfected cells compared to control silencing RNA-transfected cells was observed. The averages and standard deviations of three replicates are shown. ( d ) The difference in cell viability was compared between the negative control siRNA and IFITM3 siRNA-transfected MARC-145 cells. The graph shows the mean absorbance values read at 450 nm between the two groups. A p -value < 0.05 was considered significant.

Journal: Microorganisms

Article Title: Interferon-Induced Transmembrane Protein 3 (IFITM3) Restricts PRRSV Replication via Post-Entry Mechanisms

doi: 10.3390/microorganisms13081737

Figure Lengend Snippet: The silencing of IFITM3 slightly enhances PRRSV replication. ( a ) qRT-PCR showing the silencing of IFITM3. The averages and standard deviations of three replicates are shown. ( b ) qRT-PCR showing the viral RNA copies in control silencing and IFITM3 silencing RNA-transfected cells. The averages and standard deviations of three replicates are shown. ( c ) A slightly enhanced TCID 50 virus titer (1.22-fold increase) in IFITM3 silencing RNA-transfected cells compared to control silencing RNA-transfected cells was observed. The averages and standard deviations of three replicates are shown. ( d ) The difference in cell viability was compared between the negative control siRNA and IFITM3 siRNA-transfected MARC-145 cells. The graph shows the mean absorbance values read at 450 nm between the two groups. A p -value < 0.05 was considered significant.

Techniques: Quantitative RT-PCR, Control, Transfection, Virus, Negative Control

The IFITM3 expression level is inversely correlated to PRRSV replication efficiency. ( a ) IFN-alpha induces IFITM3 expression. Relative fold changes in IFITM3 and Mx1 gene expression in PRRSV 23,983-infected groups when compared to mock-treated groups are shown. More abundant Mx1 as compared to IFITM3 mRNA is induced by IFN-alpha. Representative data show the average and standard deviation of three replicates. ( b ) IFN-alpha inhibits PRRSV infection in a dose-dependent manner. Real-time RT-PCR shows the relative fold changes in IFITM3 and PRRSV N mRNA gene expression in IFN-alpha-treated groups at different concentrations when compared to non-treated control groups. Representative data show the average of two replicates. ( c ) The over-expression of IFITM3 results in 6.2-fold increase in the total IFITM3 protein level compared to the endogenous IFITM3 expression level. ( d ) No virus titer was detected in cells over-expressing IFITM3 while 1.9 × 10 4 TCID 50 titer was detected in cells only expressing the endogenous level of IFITM3.

Journal: Microorganisms

Article Title: Interferon-Induced Transmembrane Protein 3 (IFITM3) Restricts PRRSV Replication via Post-Entry Mechanisms

doi: 10.3390/microorganisms13081737

Figure Lengend Snippet: The IFITM3 expression level is inversely correlated to PRRSV replication efficiency. ( a ) IFN-alpha induces IFITM3 expression. Relative fold changes in IFITM3 and Mx1 gene expression in PRRSV 23,983-infected groups when compared to mock-treated groups are shown. More abundant Mx1 as compared to IFITM3 mRNA is induced by IFN-alpha. Representative data show the average and standard deviation of three replicates. ( b ) IFN-alpha inhibits PRRSV infection in a dose-dependent manner. Real-time RT-PCR shows the relative fold changes in IFITM3 and PRRSV N mRNA gene expression in IFN-alpha-treated groups at different concentrations when compared to non-treated control groups. Representative data show the average of two replicates. ( c ) The over-expression of IFITM3 results in 6.2-fold increase in the total IFITM3 protein level compared to the endogenous IFITM3 expression level. ( d ) No virus titer was detected in cells over-expressing IFITM3 while 1.9 × 10 4 TCID 50 titer was detected in cells only expressing the endogenous level of IFITM3.

Techniques: Expressing, Gene Expression, Infection, Standard Deviation, Quantitative RT-PCR, Control, Over Expression, Virus

Amphotericin B treatment only partially restores PRRSV replication in cells over-expressing IFITM3. ( a ) Immunofluorescence staining of virus-infected cells in the presence or absence of amphotericin B treatment. Images were captured at 10× magnification. ( b ) Flow cytometry analysis of virus-infected cells in the presence or absence of Amphotericin B. * indicates p < 0.05 when compared to the pQCXIP + PRRSV group.

Journal: Microorganisms

Article Title: Interferon-Induced Transmembrane Protein 3 (IFITM3) Restricts PRRSV Replication via Post-Entry Mechanisms

doi: 10.3390/microorganisms13081737

Figure Lengend Snippet: Amphotericin B treatment only partially restores PRRSV replication in cells over-expressing IFITM3. ( a ) Immunofluorescence staining of virus-infected cells in the presence or absence of amphotericin B treatment. Images were captured at 10× magnification. ( b ) Flow cytometry analysis of virus-infected cells in the presence or absence of Amphotericin B. * indicates p < 0.05 when compared to the pQCXIP + PRRSV group.

Techniques: Expressing, Immunofluorescence, Staining, Virus, Infection, Flow Cytometry

Over-expression of IFITM3 does not significantly impact virus entry. ( a ) Cells over-expressing exogenous IFITM3 contain PRRSV at 3 and 24 hpi. The colocalization of PRRSV with over-expressed IFITM3 was observed (yellow arrows). Red: IFITM3; green: PRRSV N; blue: DAPI. The images were taken at 40× magnification. ( b ) The percentage of PRRSV-positive cells was significantly lower ( p < 0.0001) in IFITM3-transfected cells at 24 hpi than at 3 hpi. Quantification was performed by counting PRRSV-positive cells and IFITM3-HA-expressing cells obtained from five different microscopic fields. * indicates p < 0.0001.

Journal: Microorganisms

Article Title: Interferon-Induced Transmembrane Protein 3 (IFITM3) Restricts PRRSV Replication via Post-Entry Mechanisms

doi: 10.3390/microorganisms13081737

Figure Lengend Snippet: Over-expression of IFITM3 does not significantly impact virus entry. ( a ) Cells over-expressing exogenous IFITM3 contain PRRSV at 3 and 24 hpi. The colocalization of PRRSV with over-expressed IFITM3 was observed (yellow arrows). Red: IFITM3; green: PRRSV N; blue: DAPI. The images were taken at 40× magnification. ( b ) The percentage of PRRSV-positive cells was significantly lower ( p < 0.0001) in IFITM3-transfected cells at 24 hpi than at 3 hpi. Quantification was performed by counting PRRSV-positive cells and IFITM3-HA-expressing cells obtained from five different microscopic fields. * indicates p < 0.0001.

Techniques: Over Expression, Virus, Expressing, Transfection

The over-expression of IFITM3 reduces the level of phosphorylated Akt by 50% compared to the vector control. A representative Western blot image shows the expression level of p-Akt. The averages and standard deviations of three replicates are shown in the graph. A significant decrease in the level of p-Akt was observed in IFITM3-over-expressing and PRRSV-infected cells compared to vector-transfected and PRRSV-infected cells ( p < 0.05).

Journal: Microorganisms

Article Title: Interferon-Induced Transmembrane Protein 3 (IFITM3) Restricts PRRSV Replication via Post-Entry Mechanisms

doi: 10.3390/microorganisms13081737

Figure Lengend Snippet: The over-expression of IFITM3 reduces the level of phosphorylated Akt by 50% compared to the vector control. A representative Western blot image shows the expression level of p-Akt. The averages and standard deviations of three replicates are shown in the graph. A significant decrease in the level of p-Akt was observed in IFITM3-over-expressing and PRRSV-infected cells compared to vector-transfected and PRRSV-infected cells ( p < 0.05).

Techniques: Over Expression, Plasmid Preparation, Control, Western Blot, Expressing, Infection, Transfection

The over-expression of IFITM3 increases the ratio of LC3-II/LC3-I by 128% compared to vector control. A representative Western blot image shows the expression level of LC3-I and LC3-II. The averages and standard deviations of three replicates are shown in the graph. * Indicates significant differences compared to mock ( p < 0.05).

Journal: Microorganisms

Article Title: Interferon-Induced Transmembrane Protein 3 (IFITM3) Restricts PRRSV Replication via Post-Entry Mechanisms

doi: 10.3390/microorganisms13081737

Figure Lengend Snippet: The over-expression of IFITM3 increases the ratio of LC3-II/LC3-I by 128% compared to vector control. A representative Western blot image shows the expression level of LC3-I and LC3-II. The averages and standard deviations of three replicates are shown in the graph. * Indicates significant differences compared to mock ( p < 0.05).

Techniques: Over Expression, Plasmid Preparation, Control, Western Blot, Expressing

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