Journal: Journal of Virology

Article Title: SARS-CoV-2 Variants of Concern Hijack IFITM2 for Efficient Replication in Human Lung Cells

doi: 10.1128/jvi.00594-22

Figure Lengend Snippet: Impact of IFITMs on replication of the Omicron VOC. (A) Schematic presentation of amino acid variations in Spike proteins of the SARS-CoV-2 Delta and Omicron VOCs investigated. Abbreviations are defined in the legend to . (B) Viral N RNA levels in the supernatant of Calu-3 cells infected with the indicated SARS-CoV-2 variants. Cells were transfected with control or IFITM-targeting siRNAs as indicated. Numbers above the bars indicate n -fold reduction compared to the viral RNA levels detected in the supernatant of Calu-3 cells treated with control siRNA. Bars in panels B and C show the mean of 3 independent experiments (±SEM), each measured in technical duplicates. (C) Quantification of intracellular viral N RNA levels in Calu-3 cells 24 h postinfection with SARS-CoV-2 (MOI of 0.05). Values were normalized to GAPDH and calculated relative to the control (set to 100%). Cells were transiently transfected with siRNA—either control (CTRL) or targeting IFITM1, -2, or -3. (D) Exemplary Western blot of IFITM and viral N protein expression in Calu-3 cells treated with control or IFITM1-, -2-, and/or -3-targeting siRNA and infected with the indicated SARS-CoV-2 variants. GAPDH was detected as a loading control. P values: *, <0.05; **, <0.01; ****, <0.0001.

Article Snippet: Proteins were stained at 1:1,000 using primary antibodies against IFITM1 (Cell Signaling, catalog no. 13126S), IFITM2 (Cell Signaling, catalog no. 13530 S), IFITM3 (Cell Signaling, catalog no. 59212S), ACE2 (rabbit polyclonal) (Abcam, catalog no. ab166755), rat anti-GAPDH (Biolegend, catalog no. 607902), and SARS CoV-2 N (Sino Biologicals, catalog no. 40588-V08B) and infrared dye-labeled secondary antibodies (Li-Cor IRDye).

Techniques: Infection, Transfection, Control, Western Blot, Expressing

Journal: Journal of Virology

Article Title: SARS-CoV-2 Variants of Concern Hijack IFITM2 for Efficient Replication in Human Lung Cells

doi: 10.1128/jvi.00594-22

Figure Lengend Snippet: Expression of ACE2 and IFITM proteins in Calu-3 and iATII cells. (A) Immunoblot of ACE2, IFITM1, IFITM2, and IFITM3 in Calu-3 and iATII cells left uninfected (lanes C) or infected with the indicated SARS-CoV-2 variants. Whole-cell lysates were stained with the indicated antibodies. An unspecific signal was observed in the Calu-3 control lane stained with the CoV-2 N antibody. (B) Flow cytometric analysis of surface ACE2 expression in Calu-3 and iATII cells.

Article Snippet: Proteins were stained at 1:1,000 using primary antibodies against IFITM1 (Cell Signaling, catalog no. 13126S), IFITM2 (Cell Signaling, catalog no. 13530 S), IFITM3 (Cell Signaling, catalog no. 59212S), ACE2 (rabbit polyclonal) (Abcam, catalog no. ab166755), rat anti-GAPDH (Biolegend, catalog no. 607902), and SARS CoV-2 N (Sino Biologicals, catalog no. 40588-V08B) and infrared dye-labeled secondary antibodies (Li-Cor IRDye).

Techniques: Expressing, Western Blot, Infection, Staining, Control

Journal: The EMBO journal

Article Title: Interferon-inducible phospholipids govern IFITM3-dependent endosomal antiviral immunity.

doi: 10.15252/embj.2022112234

Figure Lengend Snippet: Figure 7. IFITM1 and plasma membrane mutants IFITM3 inhibit SARS CoV2 entry and infection.

Article Snippet: Samples were subjected to SDS–PAGE using Bolt 4–12% Bis-Tris Plus Gels (Thermo Fisher Scientific), transferred to PVDF membrane by electroblotting, and blotted with rabbit anti-IFITM3 polyclonal Ab (1:1,000 dilution, proteintech Cat# 11714-1-AP); rabbit anti-IFITM2 polyclonal Ab (1:2,500, proteintech Cat# 12769-1-AP); mouse anti-IFITM1 monoclonal Ab (1:5,000, 2023 The Authors The EMBO Journal 42: e112234 | 2023 17 of 23 D ow nloaded from https://w w w .em bopress.org on January 13, 2024 from IP 180.252.95.111. proteintech Cat# 60074-1-Ig), rabbit anti-ACE2 (1:1,000, Cell Signaling Technology 4355S), rabbit anti-HA tag antibody—ChIP Grade (1:10,000 Abcam Cat# ab9110), mouse anti-his (1:10,000 Millipore Cat# 05-909).

Techniques: Clinical Proteomics, Membrane, Infection

Journal: Cell Reports

Article Title: Quantitative Temporal Proteomic Analysis of Vaccinia Virus Infection Reveals Regulation of Histone Deacetylases by an Interferon Antagonist

doi: 10.1016/j.celrep.2019.04.042

Figure Lengend Snippet:

Article Snippet: The following primary antibodies were used at the indicated dilution, in blocking solution: rabbit anti-IFIT1 (1:1000, cat. no. PA3-848, Thermo Fisher Scientific), mouse anti-IFITM1/2/3 (1:500, cat. no. sc-374026, Santa Cruz Biotechnology), mouse anti-COL6A2 (1:500, cat. no. sc-374566, Santa Cruz Biotechnology), mouse anti-COL1A2 (1:500, cat. no. sc-376350, Santa Cruz Biotechnology), mouse anti-TRIM5 (1:500, cat. no. SC-373864, Santa Cruz Biotechnology), mouse anti-HDAC5 (1:500, cat. no. sc-133225, Santa Cruz Biotechnology), mouse anti-HDAC1 (1:500, cat. no. sc-81598, Santa Cruz Biotechnology), mouse anti-FLAG, (1:1000, F3165, Sigma-Aldrich), mouse anti-Myc (1:1000, cat. no. 2276, Cell Signaling Technology), rat anti-α-tubulin (1:10,000, clone YL1/2, cat. no. MCA77G, Serotec), rabbit anti-C6 (1:1000, described in , mouse anti-D8 (1:1000, described in ( )).

Techniques: Virus, Western Blot, Derivative Assay, Subcloning, Recombinant, Protease Inhibitor, Electron Microscopy, Bicinchoninic Acid Protein Assay, TA Cloning, Sequencing, Mass Spectrometry, CRISPR, Disruption, Plasmid Preparation, Software, Quantitative Proteomics, Imaging, Flow Cytometry, Fluorescence, Microscopy

Journal: Scientific reports

Article Title: IFITM proteins inhibit HIV-1 protein synthesis.

doi: 10.1038/s41598-018-32785-5

Figure Lengend Snippet: Figure 7. HIV Nef can help overcome IFITM-mediated restriction of protein synthesis. (A) Level of virus production in HEK293T cells transfected with 0.5 μg expression vectors for IFITMs and 0.5 μg HIV-1 NL4-3 proviral DNA with a deletion (ΔNef) was measured by p24 ELISA 48 hours post-transfection and the data normalized, fold change of virus production compared to vector control is indicated. Differences were assessed with Student’s t tests. (B) Intracellular level of p55/p24 and IFITMs was measured by immunoblotting. (C) SupT1 cells were infected with the indicated dilutions of wild type or Nef-deleted (ΔNef) HIV-1 NL4-3 inoculum and then treated with 1 μg/ml doxycycline to induce IFITM expression post-entry and 5 μM AMD3100 to limit infections to a single round. Level of virus production was measured by p24 ELISA 72 hours post-infection. Differences were assessed with Two-way ANOVA and Bonferroni post-tests. (D) C8166 cells constitutively expressing either vector control or IFITM1 were infected with either wild type or Nef-deleted (ΔNef) HIV-1 NL4-3. Levels of virus production were measured by p24 ELISA at the indicated time-points post-infection and were normalized to the levels of virus produced from vector controls. Differences were assessed with Two-way ANOVA. (E) HEK293T cells were transfected with 0.5 μg expression vectors for IFITMs and 0.5 μg HIV-1 NL4-3 proviral DNA with either wildtype NL4-3 nef or the indicated lentiviral nef alleles. Virus production was measured by p24 ELISA 48 hours post-transfection and the data normalized. Differences were assessed with Student’s t tests. (F) HEK293T cells were transfected with ΔNef HIV-1 NL4-3 proviral DNA, expression vectors for IFITMs and an increasing proportion of HIV-1 Nef-encoding vector versus empty vector in a fixed total quantity of 1 μg. Level of virus production was measured by p24 ELISA 48 hours post- transfection, while levels of viral proteins and IFITM-FLAG expression was analyzed by (G) immunoblotting. Differences were assessed by Student’s t-test. All data show mean + SEM from 3 independent experiments and *denotes p < 0.05.

Article Snippet: The following antibodies were used to detect IFITMs: human IFITM1 (clone 5B5E2, Proteintech), IFITM2 (clone 3D5F7, Proteintech) and human IFITM3 (clone EPR5242, Novus Biologicals).

Techniques: Virus, Transfection, Expressing, Enzyme-linked Immunosorbent Assay, Plasmid Preparation, Control, Western Blot, Infection, Produced

Journal: Intractable & Rare Diseases Research

Article Title: The novel role of IFITM1-3 in myogenic differentiation of C2C12 cells

doi: 10.5582/irdr.2023.01050

Figure Lengend Snippet: Primer sequences for RT-qPCR

Article Snippet: The membranes were blocked with 5% skim milk in Tris-buffered saline containing 0.1% Tween 20 (TBST) for 2 h at room temperature, and then the membranes were incubated with primary antibodies, namely, rabbit anti-desmin (ab32362, Abcam, USA), rabbit anti-MyoD (ab203383, Abcam, USA), rabbit anti-MYF5 (ab125301, Abcam, USA), rabbit anti-IFITM1 (bs-1031R, Bioss, China), rabbit anti-IFITM2 (bs-15517R, Bioss, China), mouse anti-IFITM3 (bsm-51629M, Bioss, China), and rabbit anti-GAPDH (ab8245, Abcam, USA), at 4°C overnight.

Techniques: Sequencing

Journal: Intractable & Rare Diseases Research

Article Title: The novel role of IFITM1-3 in myogenic differentiation of C2C12 cells

doi: 10.5582/irdr.2023.01050

Figure Lengend Snippet: Increased expression of IFITM1-3 during myogenic differentiation of C2C12 myoblasts. (A) Relative expression of Ifitm1-3 during the myogenic differentiation process. (B) Western blot evaluating the protein levels of IFITM1-3. (C) Quantification of band intensity as described above is shown. The level of proteins was normalized to that of GAPDH. Statistical significance: *P < 0.05; **P < 0.01; ***P < 0.001; NS, not significant.

Article Snippet: The membranes were blocked with 5% skim milk in Tris-buffered saline containing 0.1% Tween 20 (TBST) for 2 h at room temperature, and then the membranes were incubated with primary antibodies, namely, rabbit anti-desmin (ab32362, Abcam, USA), rabbit anti-MyoD (ab203383, Abcam, USA), rabbit anti-MYF5 (ab125301, Abcam, USA), rabbit anti-IFITM1 (bs-1031R, Bioss, China), rabbit anti-IFITM2 (bs-15517R, Bioss, China), mouse anti-IFITM3 (bsm-51629M, Bioss, China), and rabbit anti-GAPDH (ab8245, Abcam, USA), at 4°C overnight.

Techniques: Expressing, Western Blot

Journal: Intractable & Rare Diseases Research

Article Title: The novel role of IFITM1-3 in myogenic differentiation of C2C12 cells

doi: 10.5582/irdr.2023.01050

Figure Lengend Snippet: Knockdown of Ifitm1, 2, and 3 by siRNAs blocks myogenic differentiation in C2C12 cells. (A) Microscopic images of Giemsa staining for C2C12 myoblasts on day 3 of myogenic differentiation after transfection with siRNAs targeting Ifitm1-3. Two different siRNAs were used for each targeting gene. Transfection without siRNAs, but with transfection reagent was set as mock. Magnification: 100×. (B) Percentage fusion on day 3 of myogenic induction and transfection with siRNAs as described above, calculated by dividing the number of nuclei within multinucleated myofibers by the total number of nuclei. NC represents the group without siRNAs and transfection reagents. (C) Downregulated protein expression of myogenin, MyoD, Myf5, and desmin after interference by siRNAs targeting Ifitm1-3. (D) Quantification of band intensity as described above is shown. The level of proteins was normalized to that of GAPDH. Statistical significance: *P< 0.05; **P < 0.01; ***P < 0.001; NS, not significant.

Article Snippet: The membranes were blocked with 5% skim milk in Tris-buffered saline containing 0.1% Tween 20 (TBST) for 2 h at room temperature, and then the membranes were incubated with primary antibodies, namely, rabbit anti-desmin (ab32362, Abcam, USA), rabbit anti-MyoD (ab203383, Abcam, USA), rabbit anti-MYF5 (ab125301, Abcam, USA), rabbit anti-IFITM1 (bs-1031R, Bioss, China), rabbit anti-IFITM2 (bs-15517R, Bioss, China), mouse anti-IFITM3 (bsm-51629M, Bioss, China), and rabbit anti-GAPDH (ab8245, Abcam, USA), at 4°C overnight.

Techniques: Staining, Transfection, Expressing

Journal: Intractable & Rare Diseases Research

Article Title: The novel role of IFITM1-3 in myogenic differentiation of C2C12 cells

doi: 10.5582/irdr.2023.01050

Figure Lengend Snippet: Identification and validation of IFITM1,3-interacting proteins. (A) Principle of co-immunoprecipitation and liquid chromatography combined with tandem mass spectrometry (LC-MS/MS). (B) The protein- protein interaction network if IFITM1,3 (overlapped) revealed by STRING analysis. A total of 84 unique homologous proteins are shown in the network. Three clusters are indicated in different colors. Cluster 1: muscle filament sliding (green color); Cluster 2: ribosome series proteins (red color); Cluster 3: regulation of mRNA metabolic process (blue color). Associations are represented by the lines. Thicker lines represent stronger associations. (C) Co-IP assays show the interaction between desmin and IFITM1, 3.

Article Snippet: The membranes were blocked with 5% skim milk in Tris-buffered saline containing 0.1% Tween 20 (TBST) for 2 h at room temperature, and then the membranes were incubated with primary antibodies, namely, rabbit anti-desmin (ab32362, Abcam, USA), rabbit anti-MyoD (ab203383, Abcam, USA), rabbit anti-MYF5 (ab125301, Abcam, USA), rabbit anti-IFITM1 (bs-1031R, Bioss, China), rabbit anti-IFITM2 (bs-15517R, Bioss, China), mouse anti-IFITM3 (bsm-51629M, Bioss, China), and rabbit anti-GAPDH (ab8245, Abcam, USA), at 4°C overnight.

Techniques: Immunoprecipitation, Liquid Chromatography, Mass Spectrometry, Liquid Chromatography with Mass Spectroscopy, Co-Immunoprecipitation Assay

Journal: Intractable & Rare Diseases Research

Article Title: The novel role of IFITM1-3 in myogenic differentiation of C2C12 cells

doi: 10.5582/irdr.2023.01050

Figure Lengend Snippet: Overlapped interacted proteins for IFITM1 and IFITM3

Article Snippet: The membranes were blocked with 5% skim milk in Tris-buffered saline containing 0.1% Tween 20 (TBST) for 2 h at room temperature, and then the membranes were incubated with primary antibodies, namely, rabbit anti-desmin (ab32362, Abcam, USA), rabbit anti-MyoD (ab203383, Abcam, USA), rabbit anti-MYF5 (ab125301, Abcam, USA), rabbit anti-IFITM1 (bs-1031R, Bioss, China), rabbit anti-IFITM2 (bs-15517R, Bioss, China), mouse anti-IFITM3 (bsm-51629M, Bioss, China), and rabbit anti-GAPDH (ab8245, Abcam, USA), at 4°C overnight.

Techniques:

Journal: Intractable & Rare Diseases Research

Article Title: The novel role of IFITM1-3 in myogenic differentiation of C2C12 cells

doi: 10.5582/irdr.2023.01050

Figure Lengend Snippet: GO and KEGG pathway enrichment analysis of 84 proteins that interact with IFITM1, 3 (overlapped). (A) KEGG classification map of differentially expressed genes. The y-axis shows the metabolic pathway. (B) Biological process (BP). (C) Cellular component (CC). (D) Molecular function (MF). The x-axis represents gene ratio = count/set size. Dot size represents the number of genes, and the color bar represents the Padj-value.

Article Snippet: The membranes were blocked with 5% skim milk in Tris-buffered saline containing 0.1% Tween 20 (TBST) for 2 h at room temperature, and then the membranes were incubated with primary antibodies, namely, rabbit anti-desmin (ab32362, Abcam, USA), rabbit anti-MyoD (ab203383, Abcam, USA), rabbit anti-MYF5 (ab125301, Abcam, USA), rabbit anti-IFITM1 (bs-1031R, Bioss, China), rabbit anti-IFITM2 (bs-15517R, Bioss, China), mouse anti-IFITM3 (bsm-51629M, Bioss, China), and rabbit anti-GAPDH (ab8245, Abcam, USA), at 4°C overnight.

Techniques:

Journal: Oncotarget

Article Title: Interferon-induced transmembrane protein 1 (IFITM1) is required for the progression of colorectal cancer

doi: 10.18632/oncotarget.13325

Figure Lengend Snippet: ( A ) IFITM1 expression was evaluated in colorectal cancer cell lines. Cell lysates were obtained from the indicated cancer cell lines, and immunoblots were performed to evaluate the expression of IFITM1. ACTIN was used as the loading control. ( B ) Proteins were quantified by Image J and expressed as the ratio of IFITM1 to ACTIN ( n = 3, * p < 0.05).

Article Snippet: The sections were incubated with anti-rabbit IFITM1 antibody (GeneTex, 1:500) for 2 hrs at room temperature, followed by incubation in enhancer for 30 mins and treatment with polymer for 1 hr at room temperature.

Techniques: Expressing, Western Blot, Control

Journal: Oncotarget

Article Title: Interferon-induced transmembrane protein 1 (IFITM1) is required for the progression of colorectal cancer

doi: 10.18632/oncotarget.13325

Figure Lengend Snippet: Colorectal cancer cell lines were infected with either control (shLacZ) virus or IFITM1 knockdown virus (642-shIFITM1 and 870-shIFITM1) and selected with puromycin. RNA and proteins were isolated to determine the expression of IFITM1. ( A ) RT-qPCR was performed to determine IFITM1 mRNA expression in SW620 cells after infection. (* p < 0.05). ( B ) Immunoblot with anti-IFITM1 antibody was conducted to analyze IFITM1 protein level, and ACTIN was used as the loading control. ( C – E ) Control (shLacZ) or IFITM1-depleted (shIFITM1) colorectal cancer cell lines were incubated for 72 hrs to determine the proliferation rate by MTT assay. Data shown are from two to three experiments (* p < 0.05, ** p < 0.001).

Article Snippet: The sections were incubated with anti-rabbit IFITM1 antibody (GeneTex, 1:500) for 2 hrs at room temperature, followed by incubation in enhancer for 30 mins and treatment with polymer for 1 hr at room temperature.

Techniques: Infection, Control, Virus, Knockdown, Isolation, Expressing, Quantitative RT-PCR, Western Blot, Incubation, MTT Assay

Journal: Oncotarget

Article Title: Interferon-induced transmembrane protein 1 (IFITM1) is required for the progression of colorectal cancer

doi: 10.18632/oncotarget.13325

Figure Lengend Snippet: Control (shLacZ) or IFITM1-depleted (shIFITM1) colorectal cancer cell lines were seeded in a matrigel-uncoated and coated transwell, followed by incubation for 18 hrs and 24 hrs for assessing migration and invasion, respectively. Cells that had migrated to the lower surface of the transwell were stained and quantified. ( A and E ) Imaging was done using an inverted microscope (magnification: 100×) and representative images are shown. Data shown (A–D: migration; E – H : invasion) are from three experiments (* p < 0.05, ** p < 0.001).

Article Snippet: The sections were incubated with anti-rabbit IFITM1 antibody (GeneTex, 1:500) for 2 hrs at room temperature, followed by incubation in enhancer for 30 mins and treatment with polymer for 1 hr at room temperature.

Techniques: Control, Incubation, Migration, Staining, Imaging, Inverted Microscopy

Journal: Oncotarget

Article Title: Interferon-induced transmembrane protein 1 (IFITM1) is required for the progression of colorectal cancer

doi: 10.18632/oncotarget.13325

Figure Lengend Snippet: ( A – D ) RNA was isolated from control (shLacZ) or IFITM1-depleted (shIFITM1) cancer cell lines (A and B: HT29; C and D: SW620) and EMT signature genes were determined by RT-qPCR (* p < 0.05, ** p < 0.001). ( E ) Cell lysates were obtained from control (shLacZ) or IFITM1-depleted (shIFITM1) HT29 cells and immunoblots were conducted with antibodies indicated. ACTIN was used as a loading control. ( F ) Supernatants obtained from control (shLacZ) or IFITM1-depleted (shIFITM1) HT29 cells were analyzed for enzymatic activity of MMP1/3, MMP2 and MMP9.

Article Snippet: The sections were incubated with anti-rabbit IFITM1 antibody (GeneTex, 1:500) for 2 hrs at room temperature, followed by incubation in enhancer for 30 mins and treatment with polymer for 1 hr at room temperature.

Techniques: Isolation, Control, Quantitative RT-PCR, Western Blot, Activity Assay

Journal: Oncotarget

Article Title: Interferon-induced transmembrane protein 1 (IFITM1) is required for the progression of colorectal cancer

doi: 10.18632/oncotarget.13325

Figure Lengend Snippet: Control (shLacZ) or IFITM1-depleted (shIFITM1) SW620 cells were treated with either control siRNA or siCAV1 for 2 days and RT-qPCR was conducted to measure the mRNA expression of IFITM1 ( A ) Caveolin-1 ( B ) and EMT signature ( D ) (* p < 0.05, ** p < 0.001). ( C ) Control (shLacZ) or IFITM1-depleted (shIFITM1) SW620 cells were treated with either control siRNA or siCAV1 and then analyzed for migration ability (C). Data shown are from three experiments (** p < 0.001).

Article Snippet: The sections were incubated with anti-rabbit IFITM1 antibody (GeneTex, 1:500) for 2 hrs at room temperature, followed by incubation in enhancer for 30 mins and treatment with polymer for 1 hr at room temperature.

Techniques: Control, Quantitative RT-PCR, Expressing, Migration

Journal: Oncotarget

Article Title: Interferon-induced transmembrane protein 1 (IFITM1) is required for the progression of colorectal cancer

doi: 10.18632/oncotarget.13325

Figure Lengend Snippet: ( A and B ) SW620 cells were transiently transfected with either control (vector) or IFITM1 expression construct (IFITM1 OE). RNA and proteins were isolated to examine the expression of IFITM1. (A) RT-qPCR was performed to determine IFITM1 mRNA expression in SW620 cells after transfection (** p < 0.001). (B) IFITM1 protein level was analyzed by immunoblot using anti-IFITM1 antibody, and ACTIN was used as the loading control. ( C ) SW620 cells transfected with control (vector) or IFITM1-expression construct (IFITM1 OE) were incubated for 72 hrs to determine the proliferation by MTT assay. Data shown are from three experiments (** p < 0.001). ( D and E ) SW620 cells transfected with control (vector) or IFITM1-expression construct (IFITM1 OE) were seeded in a transwell for assessing migration. Cells were incubated for 18 hrs, stained, and quantified. (D) Representative images are shown. ( E ) Data shown are from three experiments (* p < 0.05). ( F and G ) SW620 cells were transiently transfected with either Vector alone, IFITM1-overexpressing (IFITM1 OE) and Vector, or IFITM1 OE and CAV1-overexpressing (CAV1 OE) constructs. RNA was isolated from the transduced cells, and CAV1 (F) and EMT signature (G) were quantified by RT-qPCR (* p < 0.05, ** p < 0.001).

Article Snippet: The sections were incubated with anti-rabbit IFITM1 antibody (GeneTex, 1:500) for 2 hrs at room temperature, followed by incubation in enhancer for 30 mins and treatment with polymer for 1 hr at room temperature.

Techniques: Transfection, Control, Plasmid Preparation, Expressing, Construct, Isolation, Quantitative RT-PCR, Western Blot, Incubation, MTT Assay, Migration, Staining

Journal: Oncotarget

Article Title: Interferon-induced transmembrane protein 1 (IFITM1) is required for the progression of colorectal cancer

doi: 10.18632/oncotarget.13325

Figure Lengend Snippet: ( A and B ) RNA was isolated from formalin-fixed and paraffin-embedded (FFPE) tissues and IFITM1 was quantified by RT-qPCR. (N: normal; T: tumor; META: metastasis samples). Individual samples (A) and average of each group of samples (B) are shown (* p < 0.05, ** p < 0.001). ( C – E ) 232 colorectal cancer samples were stained with IFITM1 antibody and graded based on both staining intensity and staining frequency. (C and D) Representative images of tissue microarray and immunohistochemistry are shown. (E) Survival rate was determined based on the expression of IFITM1 expression using Kaplan-Meier method ( p < 0.05).

Article Snippet: The sections were incubated with anti-rabbit IFITM1 antibody (GeneTex, 1:500) for 2 hrs at room temperature, followed by incubation in enhancer for 30 mins and treatment with polymer for 1 hr at room temperature.

Techniques: Isolation, Quantitative RT-PCR, Staining, Microarray, Immunohistochemistry, Expressing

Journal: Oncotarget

Article Title: Interferon-induced transmembrane protein 1 (IFITM1) is required for the progression of colorectal cancer

doi: 10.18632/oncotarget.13325

Figure Lengend Snippet: Association of IFITM1 expression with clinicopathological factors by logistic regression analysis

Article Snippet: The sections were incubated with anti-rabbit IFITM1 antibody (GeneTex, 1:500) for 2 hrs at room temperature, followed by incubation in enhancer for 30 mins and treatment with polymer for 1 hr at room temperature.

Techniques: Expressing

Journal: Oncogenesis

Article Title: A mesenchymal glioma stem cell profile is related to clinical outcome

doi: 10.1038/oncsis.2014.5

Figure Lengend Snippet: Hierarchical clustering analysis of GSCs. ( a ) The unsupervised clustering using Ward's linkage method and Euclidean distance individualized H9 HNSCs, H1-H9-ESCs and two groups of GSCs (GSCX-1 and GSCX-2). Note that culture conditions of H9 HNSC do not influence the expression of pluripotency gene samples H9 HNSC-1 and H9 HNSC-2 samples. ( b ) Box plot representation of COL1A1 and IFITM1 expression levels between GSC groups GSCX-1 and GSCX-2. The charts show the log 10 expression of relative quantification (RQ) values normalized to the expression of H9 HNSC cell line. ( c ) Box plot representation of COL1A1 and IFITM1 expression levels between bulk tumor groups TX-1 and TX-2. The top edge of the boxes represents the 75th percentile, the bottom edge, the 25th percentile and × the mean. The range is shown as vertical edge.

Article Snippet: Antibodies used in this study were goat anti-IFITM1 antibody (R&D Systems Europe, Lille, France), rabbit anti-COL1A1 antibody (Abcam, Paris, France) and mouse β-actin antibody (Abcam).

Techniques: Expressing, Quantitative Proteomics

Journal: Oncogenesis

Article Title: A mesenchymal glioma stem cell profile is related to clinical outcome

doi: 10.1038/oncsis.2014.5

Figure Lengend Snippet: Knockdown efficiency of COL1A1 and IFITM1 and effect on cell invasion. ( a ) COL1A1 and IFITM1 mRNA expression was monitored by quantitative reverse transcriptase–PCR in GSC-3 and GSC-9 ( n =2). Significant inhibition of 90% of mRNA expression was observed. GADPH was used as internal control. * P <0.05. Each value is expressed as mean±s.e.m. ( b ) Western blot analysis showed diminution of IFITM1 and COL1A1 proteins expression, β-actin was used as loading control. ( c ) Densitometry of protein level. The bar graph shows relative IFITM1 and COL1A1 protein expression levels expressed as percentage of IFITM1 and COL1A1 expression in control cells ( n =3). We observed >80% inhibition for both genes in GSC-3 and GSC-9 cells, as compared with control cells. ( d ) Representative fields of Boyden cell invasion assay. After 3 days of invasion, cells that migrated across the membrane were fixed and stained with hematoxylin/eosin and were counted in one hundred different fields. The invaded cells were photographed under the microscope at × 100 magnification. ( e ) A significant reduction of invasion is observed in both GSC-3 and GSC-9 cells lines with sh-IFITM1 as well as for sh-COL1A1. The results represent a percentage of invaded cells. Data are presented as mean±s.e.m. from three independent experiments. * P <0.05, ** P <0.01.

Article Snippet: Antibodies used in this study were goat anti-IFITM1 antibody (R&D Systems Europe, Lille, France), rabbit anti-COL1A1 antibody (Abcam, Paris, France) and mouse β-actin antibody (Abcam).

Techniques: Knockdown, Expressing, Reverse Transcription, Inhibition, Control, Western Blot, Invasion Assay, Membrane, Staining, Microscopy

Journal: Oncogenesis

Article Title: A mesenchymal glioma stem cell profile is related to clinical outcome

doi: 10.1038/oncsis.2014.5

Figure Lengend Snippet: Effects of COL1A1 and IFITM1 inhibition on cell proliferation and on neurosphere-initiating capacity. ( a ) Cell proliferation was measured by using a MTS assay. Each experiment was carried out in replicates of six and repeated twice and expressed as mean±s.d. ( b ) Inhibition of COL1A1 resulted in a significant (* P <0.05) decrease of neurosphere-initiating cell (NS-IC) frequency whereas inhibition of IFITM1 expression only slightly modified the neurosphere-initiating capacity. Final cell dilutions yielding 37% of negative wells correspond to the dilution at which there is one NS-ICs per well. Each experiment was performed in duplicate and repeated twice and expressed as mean±s.d.

Article Snippet: Antibodies used in this study were goat anti-IFITM1 antibody (R&D Systems Europe, Lille, France), rabbit anti-COL1A1 antibody (Abcam, Paris, France) and mouse β-actin antibody (Abcam).

Techniques: Inhibition, MTS Assay, Expressing, Modification

Journal: Oncogenesis

Article Title: A mesenchymal glioma stem cell profile is related to clinical outcome

doi: 10.1038/oncsis.2014.5

Figure Lengend Snippet: COL1A1 and IFITM1 expression levels in an independent cohort of 30 patients' tumors. ( a ) Box plot representation of the mRNA expression levels. The charts show the log 10 expression of relative quantification (RQ) values normalized to the expression of H9 NSC cell line. The top edge of the boxes represents the 75th percentile, the bottom edge, the 25th percentile and × the mean. The range is shown as vertical edge. ( b ) Kaplan–Meier analysis for overall survival of an independent cohort of 30 GBM patients. Survival distribution of patients with low (group A; n =13) versus high (group B; n =17) expression of COL1A1 and IFITM1 ( P <10 −4 ).

Article Snippet: Antibodies used in this study were goat anti-IFITM1 antibody (R&D Systems Europe, Lille, France), rabbit anti-COL1A1 antibody (Abcam, Paris, France) and mouse β-actin antibody (Abcam).

Techniques: Expressing, Quantitative Proteomics