abcf1 gene  (Santa Cruz Biotechnology)

Journal: bioRxiv

Article Title: An ATP-Binding Cassette Transporter Gene Links Innate and Adaptive Immune Responses

doi: 10.1101/2023.12.31.573785

Figure Lengend Snippet: ABCF1 promoter activity in CD8+ T cells from the thymus and the spleen. Spleens and thymi were dissected from threeABCF1+/- (top panels) and three of their wild-type littermate controls (bottom panels). Cells were labelled with the αnti-CD8 antibody and αnti-CD4 antibody. We depict the staining of two ABCF1+/- and one wild-type littermate controls. Data is representative of two separate experiments.

Article Snippet: Specifically, knockdown of the ABCF1 gene was accomplished using Mouse Abcf1 siRNA obtained from Santa Cruz Biotechnology (catalog number sc-140760), consistently resulting in a reduction of 79-81% compared to control samples treated with scrambled siRNA.

Techniques: Activity Assay, Staining

Journal: bioRxiv

Article Title: An ATP-Binding Cassette Transporter Gene Links Innate and Adaptive Immune Responses

doi: 10.1101/2023.12.31.573785

Figure Lengend Snippet: ABCF1 promoter activity in CD8+ T cells from the thymus and the spleen. Spleens and thymi were dissected out of three ABCF1+/- (top panels) and three of their wild-type littermate controls (bottom panels). Cells were labelled with the PE-αCD8 antibody and then stained with fluorescein by heat shock. Data is representative of two separate experiments.

Article Snippet: Specifically, knockdown of the ABCF1 gene was accomplished using Mouse Abcf1 siRNA obtained from Santa Cruz Biotechnology (catalog number sc-140760), consistently resulting in a reduction of 79-81% compared to control samples treated with scrambled siRNA.

Techniques: Activity Assay, Staining

Journal: bioRxiv

Article Title: An ATP-Binding Cassette Transporter Gene Links Innate and Adaptive Immune Responses

doi: 10.1101/2023.12.31.573785

Figure Lengend Snippet: ABCF1 +/- mice produce fewer IFN γ positive CD8+ T lymphocytes after infection with VSV. ABCF1+/- splenocytes showed fewer IFNγ positive CD8+ T lymphocytes. Figure shown represents the results of one experiment, which was later repeated three times with similar results.

Article Snippet: Specifically, knockdown of the ABCF1 gene was accomplished using Mouse Abcf1 siRNA obtained from Santa Cruz Biotechnology (catalog number sc-140760), consistently resulting in a reduction of 79-81% compared to control samples treated with scrambled siRNA.

Techniques: Infection

Journal: bioRxiv

Article Title: An ATP-Binding Cassette Transporter Gene Links Innate and Adaptive Immune Responses

doi: 10.1101/2023.12.31.573785

Figure Lengend Snippet: ABCF1+/- CTLs were as efficient as ABCF1 +/+ at killing of chromium. loaded VSV specific target lymphocytes. CTL activity was measured by the release of 51 Cr into the supernatant. Figure shown represents the results of one experiment, which was later repeated three times with similar results.

Article Snippet: Specifically, knockdown of the ABCF1 gene was accomplished using Mouse Abcf1 siRNA obtained from Santa Cruz Biotechnology (catalog number sc-140760), consistently resulting in a reduction of 79-81% compared to control samples treated with scrambled siRNA.

Techniques: Activity Assay

Journal: bioRxiv

Article Title: An ATP-Binding Cassette Transporter Gene Links Innate and Adaptive Immune Responses

doi: 10.1101/2023.12.31.573785

Figure Lengend Snippet: ABCF1 is essential for the production of ISG (Interferon-Stimulated Gene) specific cytokines expression of INF- α and β after Poly(I:C) stimulation. A bar graph illustrating the fold change in the levels of INF-α and β cytokines detected in the cell culture supernatants. These findings provide insights into the impact of Abcf1 -specific siRNA treatment, both with and without Poly(I:C), on the secretion of cytokines and chemokines as well as the activation of key signaling pathways and transcription factors within BMDMs. Expression levels of the protein was assessed by western blotting analysis.

Article Snippet: Specifically, knockdown of the ABCF1 gene was accomplished using Mouse Abcf1 siRNA obtained from Santa Cruz Biotechnology (catalog number sc-140760), consistently resulting in a reduction of 79-81% compared to control samples treated with scrambled siRNA.

Techniques: Expressing, Cell Culture, Activation Assay, Protein-Protein interactions, Western Blot

Journal: bioRxiv

Article Title: An ATP-Binding Cassette Transporter Gene Links Innate and Adaptive Immune Responses

doi: 10.1101/2023.12.31.573785

Figure Lengend Snippet: ABCF1 plays a role in regulating the phosphorylation levels of NF- κ B p65 and IRF3. A) Cytoplasmic and nuclear fractions obtained from BMDMs under different conditions were subjected to western blot analysis to assess the levels of transcription factors associated with Poly(I:C) stimulation confirming our hypothesis; Western Blot band sizes are as follows: anti- p65 (65 kD); anti p-p65 (65 kD); anti IRF3 (47 kD); anti p-IRF3 (36 kD); anti ABCF1 (110 kD); anti GAPDH (36 kD); anti histone H3 (15 kD); Dimerized-IRF3 (73 kD); IRF3 (36 kD) B) Whole-cell lysates were separated using native PAGE gel electrophoresis, and the analysis focused on examining the dimerization of IRF3, a transcription factor. Expression levels of the protein was assessed by western blotting analysis. Fold change calculations, as specified in the materials and methods section, were employed to quantify changes in the data. The bar graphs displayed in the figures represent the mean values along with standard deviations. The data presented here is representative of three separate experiments.

Article Snippet: Specifically, knockdown of the ABCF1 gene was accomplished using Mouse Abcf1 siRNA obtained from Santa Cruz Biotechnology (catalog number sc-140760), consistently resulting in a reduction of 79-81% compared to control samples treated with scrambled siRNA.

Techniques: Phospho-proteomics, Western Blot, Clear Native PAGE, Nucleic Acid Electrophoresis, Expressing

Journal: bioRxiv

Article Title: An ATP-Binding Cassette Transporter Gene Links Innate and Adaptive Immune Responses

doi: 10.1101/2023.12.31.573785

Figure Lengend Snippet: ABCF1 Parallels OAS1a Expression and Controls 2-5A Activity A) The immunoprecipitates were immunoblotted with anti-ABCF1 and anti-OAS1a antibodies. B) ABCF1 was over-expressed (Over Exp.) in BMDMs, an equal amount of whole cell lysates was immunoprecipitated with anti-ABCF1 antibody and the immunoprecipitates were detected with anti-ABCF1 and anti-OAS1a antibodies using western blots. C) Protein levels of OAS1a, ABCE1 and RNase L were analyzed from whole cell lysates from Abcf1 siRNA-treated BMDMs in presence or absence of Poly(I:C). Expression levels of the protein was assessed by western blotting analysis. D) Increased pyrophosphate production (a readout of 2-5A activity) was measured in scrambled and Abcf1 siRNA treated BMDMs in presence and absence of recombinant OAS1. E) Total RNA was extracted from Abcf1 siRNA treated BMDMs in presence or absence of Poly(I:C) and was run on a Agilent 2100 Bioanalyzer. 28S and 18S RNA have been shown and RIN score was calculated. Bars indicate the mean ±standard deviation. The data is representative of 3 separate experiments. The expression of ABCF1 follows a pattern that parallels the expression of OAS1a, and it also exerts control over the activity of 2-5A. This suggests a regulatory relationship where ABCF1’s expression correlates with OAS1a expression and influences the activity of 2-5A.

Article Snippet: Specifically, knockdown of the ABCF1 gene was accomplished using Mouse Abcf1 siRNA obtained from Santa Cruz Biotechnology (catalog number sc-140760), consistently resulting in a reduction of 79-81% compared to control samples treated with scrambled siRNA.

Techniques: Expressing, Activity Assay, Immunoprecipitation, Western Blot, Recombinant, Standard Deviation, Control

Journal: bioRxiv

Article Title: An ATP-Binding Cassette Transporter Gene Links Innate and Adaptive Immune Responses

doi: 10.1101/2023.12.31.573785

Figure Lengend Snippet: ABCF1 promoter activity in CD8+ T cells from the thymus and the spleen. Spleens and thymi were dissected from threeABCF1+/- (top panels) and three of their wild-type littermate controls (bottom panels). Cells were labelled with the αnti-CD8 antibody and αnti-CD4 antibody. We depict the staining of two ABCF1+/- and one wild-type littermate controls. Data is representative of two separate experiments.

Article Snippet: Specifically, knockdown of the Abcf1 gene utilized Mouse Abcf1 siRNA from Santa Cruz Biotechnology (catalog number sc-140760), resulting in a consistent reduction of 79-81% compared to control samples treated with scrambled siRNA.

Techniques: Activity Assay, Staining

Journal: bioRxiv

Article Title: An ATP-Binding Cassette Transporter Gene Links Innate and Adaptive Immune Responses

doi: 10.1101/2023.12.31.573785

Figure Lengend Snippet: ABCF1 promoter activity in CD8+ T cells from the thymus and the spleen. Spleens and thymi were dissected out of three ABCF1+/- (top panels) and three of their wild-type littermate controls (bottom panels). Cells were labelled with the PE-αCD8 antibody and then stained with fluorescein by heat shock. Data is representative of two separate experiments.

Article Snippet: Specifically, knockdown of the Abcf1 gene utilized Mouse Abcf1 siRNA from Santa Cruz Biotechnology (catalog number sc-140760), resulting in a consistent reduction of 79-81% compared to control samples treated with scrambled siRNA.

Techniques: Activity Assay, Staining

Journal: bioRxiv

Article Title: An ATP-Binding Cassette Transporter Gene Links Innate and Adaptive Immune Responses

doi: 10.1101/2023.12.31.573785

Figure Lengend Snippet: ABCF1 +/- mice produce fewer IFN γ positive CD8+ T lymphocytes after infection with VSV. ABCF1+/- splenocytes showed fewer IFNγ positive CD8+ T lymphocytes. Figure shown represents the results of one experiment, which was later repeated three times with similar results.

Article Snippet: Specifically, knockdown of the Abcf1 gene utilized Mouse Abcf1 siRNA from Santa Cruz Biotechnology (catalog number sc-140760), resulting in a consistent reduction of 79-81% compared to control samples treated with scrambled siRNA.

Techniques: Infection

Journal: bioRxiv

Article Title: An ATP-Binding Cassette Transporter Gene Links Innate and Adaptive Immune Responses

doi: 10.1101/2023.12.31.573785

Figure Lengend Snippet: ABCF1+/- CTLs were as efficient as ABCF1 +/+ at killing of chromium. loaded VSV specific target lymphocytes. CTL activity was measured by the release of 51 Cr into the supernatant. Figure shown represents the results of one experiment, which was later repeated three times with similar results.

Article Snippet: Specifically, knockdown of the Abcf1 gene utilized Mouse Abcf1 siRNA from Santa Cruz Biotechnology (catalog number sc-140760), resulting in a consistent reduction of 79-81% compared to control samples treated with scrambled siRNA.

Techniques: Activity Assay

Journal: bioRxiv

Article Title: An ATP-Binding Cassette Transporter Gene Links Innate and Adaptive Immune Responses

doi: 10.1101/2023.12.31.573785

Figure Lengend Snippet: ABCF1 is essential for the production of ISG (Interferon-Stimulated Gene) specific cytokines expression of INF- α and β after Poly(I:C) stimulation. A bar graph illustrating the fold change in the levels of INF-α and β cytokines detected in the cell culture supernatants. These findings provide insights into the impact of Abcf1 -specific siRNA treatment, both with and without Poly(I:C), on the secretion of cytokines and chemokines as well as the activation of key signaling pathways and transcription factors within BMDMs. Expression levels of the protein was assessed by western blotting analysis.

Article Snippet: Specifically, knockdown of the Abcf1 gene utilized Mouse Abcf1 siRNA from Santa Cruz Biotechnology (catalog number sc-140760), resulting in a consistent reduction of 79-81% compared to control samples treated with scrambled siRNA.

Techniques: Expressing, Cell Culture, Activation Assay, Protein-Protein interactions, Western Blot

Journal: bioRxiv

Article Title: An ATP-Binding Cassette Transporter Gene Links Innate and Adaptive Immune Responses

doi: 10.1101/2023.12.31.573785

Figure Lengend Snippet: ABCF1 plays a role in regulating the phosphorylation levels of NF- κ B p65 and IRF3. A) Cytoplasmic and nuclear fractions obtained from BMDMs under different conditions were subjected to western blot analysis to assess the levels of transcription factors associated with Poly(I:C) stimulation confirming our hypothesis; Western Blot band sizes are as follows: anti- p65 (65 kD); anti p-p65 (65 kD); anti IRF3 (47 kD); anti p-IRF3 (36 kD); anti ABCF1 (110 kD); anti GAPDH (36 kD); anti histone H3 (15 kD); Dimerized-IRF3 (73 kD); IRF3 (36 kD) B) Whole-cell lysates were separated using native PAGE gel electrophoresis, and the analysis focused on examining the dimerization of IRF3, a transcription factor. Expression levels of the protein was assessed by western blotting analysis. Fold change calculations, as specified in the materials and methods section, were employed to quantify changes in the data. The bar graphs displayed in the figures represent the mean values along with standard deviations. The data presented here is representative of three separate experiments.

Article Snippet: Specifically, knockdown of the Abcf1 gene utilized Mouse Abcf1 siRNA from Santa Cruz Biotechnology (catalog number sc-140760), resulting in a consistent reduction of 79-81% compared to control samples treated with scrambled siRNA.

Techniques: Phospho-proteomics, Western Blot, Clear Native PAGE, Nucleic Acid Electrophoresis, Expressing

Journal: bioRxiv

Article Title: An ATP-Binding Cassette Transporter Gene Links Innate and Adaptive Immune Responses

doi: 10.1101/2023.12.31.573785

Figure Lengend Snippet: ABCF1 Parallels OAS1a Expression and Controls 2-5A Activity A) The immunoprecipitates were immunoblotted with anti-ABCF1 and anti-OAS1a antibodies. B) ABCF1 was over-expressed (Over Exp.) in BMDMs, an equal amount of whole cell lysates was immunoprecipitated with anti-ABCF1 antibody and the immunoprecipitates were detected with anti-ABCF1 and anti-OAS1a antibodies using western blots. C) Protein levels of OAS1a, ABCE1 and RNase L were analyzed from whole cell lysates from Abcf1 siRNA-treated BMDMs in presence or absence of Poly(I:C). Expression levels of the protein was assessed by western blotting analysis. D) Increased pyrophosphate production (a readout of 2-5A activity) was measured in scrambled and Abcf1 siRNA treated BMDMs in presence and absence of recombinant OAS1. E) Total RNA was extracted from Abcf1 siRNA treated BMDMs in presence or absence of Poly(I:C) and was run on a Agilent 2100 Bioanalyzer. 28S and 18S RNA have been shown and RIN score was calculated. Bars indicate the mean ±standard deviation. The data is representative of 3 separate experiments. The expression of ABCF1 follows a pattern that parallels the expression of OAS1a, and it also exerts control over the activity of 2-5A. This suggests a regulatory relationship where ABCF1’s expression correlates with OAS1a expression and influences the activity of 2-5A.

Article Snippet: Specifically, knockdown of the Abcf1 gene utilized Mouse Abcf1 siRNA from Santa Cruz Biotechnology (catalog number sc-140760), resulting in a consistent reduction of 79-81% compared to control samples treated with scrambled siRNA.

Techniques: Expressing, Activity Assay, Immunoprecipitation, Western Blot, Recombinant, Standard Deviation, Control

Journal: bioRxiv

Article Title: Phagocytosis in macrophages is regulated by the ATP-binding cassette family gene ABCF1

doi: 10.1101/2023.09.05.556419

Figure Lengend Snippet: A) In bone marrow-derived macrophages (BMDMs), ABCF1 was manipulated through either overexpression (Over Exp.) or treatment with specific siRNA. These interventions were carried out both in the presence and absence of lipopolysaccharide (LPS) stimulation (at a concentration of 100 ng/ml for 24 hours). The resulting heat map illustrates the fold change in various cytokines and chemokines detected in the cell culture supernatants. B) Similarly, in BMDMs, ABCF1 was modulated using Abcf1-specific siRNA, and these cells were exposed to LPS treatment. The heat map presented here showcases the fold change in various Mitogen-Activated Protein Kinase (MAPK) and Interferon-Stimulated Gene (ISG)-specific phosphoproteins and transcription factors detected in whole-cell lysates (WCL).

Article Snippet: Specifically, ABCF1 gene knockdown was accomplished using Mouse Abcf1 siRNA from Santa Cruz Biotechnology (catalog number sc-140760), resulting in a consistent reduction of 79-81% compared to control samples treated with scrambled siRNA.

Techniques: Derivative Assay, Over Expression, Concentration Assay, Cell Culture

Journal: bioRxiv

Article Title: Phagocytosis in macrophages is regulated by the ATP-binding cassette family gene ABCF1

doi: 10.1101/2023.09.05.556419

Figure Lengend Snippet: A) The absorbance value associated with phagocytosis was assessed in bone marrow-derived macrophages (BMDMs) that were subjected to ABCF1-specific siRNA treatment. These BMDMs were incubated with fluorescently labeled IgG-coated latex beads for a duration of 2 hours, both in the presence and absence of lipopolysaccharide (LPS). Additionally, ABCF1-specific siRNA-treated BMDMs were incubated with fluorescently labeled E. coli for 2 hours. The results provide insights into the impact of ABCF1 modulation on phagocytosis; B ) The levels of CD16 and CD64 were analyzed using flow cytometry in treated BMDMs. The bar graph depicts the change in mean fluorescence intensity (MFI), and error bars indicate the standard deviation. C ) The levels of CD32 were analyzed using flow cytometry in treated BMDMs. The bar graph depicts the change in mean fluorescence intensity (MFI), and error bars indicate the standard deviation. This analysis sheds light on how ABCF1 manipulation influences CD32 expression; C ) In BMDMs, ABCF1 was either overexpressed or not, and these cells were exposed to LPS. Whole-cell lysates (WCL) were immunoprecipitated with an anti-CD32 antibody. The immunoprecipitates were subsequently subjected to immunoblotting using anti-CD32 and anti-phosphotyrosine antibodies. This experimental approach allows the investigation of ABCF1’s role in CD32-associated signaling events; D ) The heat map represents the fold change in phosphorylation levels of Src Family Kinases (SFKs) detected in whole-cell lysates (WCL) from ABCF1-specific siRNA-treated and LPS-stimulated BMDMs. This analysis provides insights into how ABCF1 modulation affects SFK phosphorylation levels in this context.

Article Snippet: Specifically, ABCF1 gene knockdown was accomplished using Mouse Abcf1 siRNA from Santa Cruz Biotechnology (catalog number sc-140760), resulting in a consistent reduction of 79-81% compared to control samples treated with scrambled siRNA.

Techniques: Derivative Assay, Incubation, Labeling, Flow Cytometry, Fluorescence, Standard Deviation, Expressing, Immunoprecipitation, Western Blot, Phospho-proteomics

Journal: bioRxiv

Article Title: Phagocytosis in macrophages is regulated by the ATP-binding cassette family gene ABCF1

doi: 10.1101/2023.09.05.556419

Figure Lengend Snippet: ABCF1 plays a crucial role in orchestrating the intricate process of phagocytosis, particularly in the engulfment of E. coli particles mediated by Fcγ receptors (FcγRs). The sequence of events involving ABCF1 in this process unfolds as follows: 1 ) ABCF1 is essential for the phosphorylation of Immunoreceptor Tyrosine-Based Activation Motifs (ITAMs) by Src Family Kinases (SFKs) such as SRC, LYN, or FGR. This initial phosphorylation event is the trigger for FcγR-mediated phagocytosis of E. coli particles; 2 ) Following ITAM phosphorylation, ABCF1 takes on the role of targeting Spleen Tyrosine Kinase (SYK) for K63-linked polyubiquitination. This ubiquitination event is crucial for subsequent signaling events; 3 ) SYK, once polyubiquitinated, undergoes phosphorylation mediated by SFKs, such as FYN and other members of the SFK family. These phosphorylation events initiate downstream signaling pathways; 4 ) Phosphorylated SYK sets off a cascade of events, including the phosphorylation of Phospholipase C Gamma 2 (PLCγ2), FYN, and other SFKs. These signaling molecules play critical roles in actin polymerization and the formation of the phagocytic cup, a structural prerequisite for successful phagocytosis; 5 ) The downstream consequences of this signaling cascade include elevated phosphorylation levels of Signal Transducer and Activator of Transcription (STAT) proteins, particularly STAT 2, 3, and 6. These phosphorylated STAT proteins contribute to an increase in the production of anti-inflammatory cytokines. In summary, ABCF1’s involvement in phagocytosis of E. coli particles revolves around its role in initiating and coordinating a series of signaling events that ultimately lead to successful engulfment and the production of anti-inflammatory cytokines.

Article Snippet: Specifically, ABCF1 gene knockdown was accomplished using Mouse Abcf1 siRNA from Santa Cruz Biotechnology (catalog number sc-140760), resulting in a consistent reduction of 79-81% compared to control samples treated with scrambled siRNA.

Techniques: Sequencing, Phospho-proteomics, Activation Assay, Ubiquitin Proteomics, Protein-Protein interactions