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cpsf7  (Bethyl)


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    Bethyl cpsf7
    Loss of CPSF6 expression globally shortens the 3’UTR of cellular mRNAs. ( A ) A549 WT, NT#H1, CPSF6-KO#B4, CPSF6-KO#B7, and CPSF6-KO#C8 cells were lysed, and proteins were analyzed by Nu-PAGE, followed by western blot using anti-CPSF6, anti-CPSF5, <t>anti-CPSF7,</t> and anti-GAPDH antibodies. Experiments were repeated at least three times, and a representative image is shown. Graphs show the average densitometry quantification of three replicates with standard deviation. Significance was determined using ANOVA multiple comparisons tests; *** P < 0.001; ns, not significant. ( B ) Total RNA from CPSF6-KO and WT A549 cells was prepared, and polyadenylated transcripts were identified by PAC-seq, followed by APA analysis using PolyAMiner. ( C ) A549 WT, NT#H1, CPSF6-KO#B4, CPSF6-KO#B7, and CPSF6-KO#C8 cells were infected with increasing amounts of HIV-1-GFP for 24 or 48 h. Infection was assessed as the percentage of GFP-positive cells by flow cytometry. ( D ) WT and CPSF6-KO#B7 A549 cells were infected with HIV-1-GFP at an MOI of 2 for 48 h. Cells were fixed, permeabilized, and stained using the following antibodies: (i) anti-CPSF5 (red) and anti-CPSF6 (green); (ii) anti-SC-35 (red) and anti-CPSF6 (green); (iii) anti-LEDGF/p75 (red) and anti-CPSF6 (green); and (iv) anti-LEDGF/p75 (red) and anti-SC35 (green). Nuclei were stained with DAPI (blue). Scale bar, 10 µm. ( E ) Percentage of A549 cells containing CPSF6, CPSF5, or CPSF7 in nuclear speckles (condensates) upon HIV-1 infection (average of three independent experiments with standard deviation). Cells containing CPSF6, CPSF5, or CPSF7 in nuclear speckles were determined by visual examination of 200 cells. Significance was determined using unpaired t -test; *** P < 0.001; ns, not significant. APA, alternative polyadenylation; CPSF5, cleavage and polyadenylation specificity factor subunit 5; CPSF6, cleavage and polyadenylation specificity factor subunit 6; CPSF7, cleavage and polyadenylation specificity factor subunit 7; DAPI, 4’,6-diamidino-2-phenylindole; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; KO, knockout; LEDGF/p75, lens epithelium-derived growth factor; NT, non-targeting; PAC-Seq, Poly(A)-Click-Sequencing; UTR, untranslated region; WT, wild-type; hpi, hours post-infection.
    Cpsf7, supplied by Bethyl, used in various techniques. Bioz Stars score: 93/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cpsf7/product/Bethyl
    Average 93 stars, based on 16 article reviews
    cpsf7 - by Bioz Stars, 2026-04
    93/100 stars

    Images

    1) Product Images from "HIV-1 infection regulates gene expression by altering alternative polyadenylation correlated with CPSF6 and CPSF5 redistribution"

    Article Title: HIV-1 infection regulates gene expression by altering alternative polyadenylation correlated with CPSF6 and CPSF5 redistribution

    Journal: mBio

    doi: 10.1128/mbio.02865-25

    Loss of CPSF6 expression globally shortens the 3’UTR of cellular mRNAs. ( A ) A549 WT, NT#H1, CPSF6-KO#B4, CPSF6-KO#B7, and CPSF6-KO#C8 cells were lysed, and proteins were analyzed by Nu-PAGE, followed by western blot using anti-CPSF6, anti-CPSF5, anti-CPSF7, and anti-GAPDH antibodies. Experiments were repeated at least three times, and a representative image is shown. Graphs show the average densitometry quantification of three replicates with standard deviation. Significance was determined using ANOVA multiple comparisons tests; *** P < 0.001; ns, not significant. ( B ) Total RNA from CPSF6-KO and WT A549 cells was prepared, and polyadenylated transcripts were identified by PAC-seq, followed by APA analysis using PolyAMiner. ( C ) A549 WT, NT#H1, CPSF6-KO#B4, CPSF6-KO#B7, and CPSF6-KO#C8 cells were infected with increasing amounts of HIV-1-GFP for 24 or 48 h. Infection was assessed as the percentage of GFP-positive cells by flow cytometry. ( D ) WT and CPSF6-KO#B7 A549 cells were infected with HIV-1-GFP at an MOI of 2 for 48 h. Cells were fixed, permeabilized, and stained using the following antibodies: (i) anti-CPSF5 (red) and anti-CPSF6 (green); (ii) anti-SC-35 (red) and anti-CPSF6 (green); (iii) anti-LEDGF/p75 (red) and anti-CPSF6 (green); and (iv) anti-LEDGF/p75 (red) and anti-SC35 (green). Nuclei were stained with DAPI (blue). Scale bar, 10 µm. ( E ) Percentage of A549 cells containing CPSF6, CPSF5, or CPSF7 in nuclear speckles (condensates) upon HIV-1 infection (average of three independent experiments with standard deviation). Cells containing CPSF6, CPSF5, or CPSF7 in nuclear speckles were determined by visual examination of 200 cells. Significance was determined using unpaired t -test; *** P < 0.001; ns, not significant. APA, alternative polyadenylation; CPSF5, cleavage and polyadenylation specificity factor subunit 5; CPSF6, cleavage and polyadenylation specificity factor subunit 6; CPSF7, cleavage and polyadenylation specificity factor subunit 7; DAPI, 4’,6-diamidino-2-phenylindole; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; KO, knockout; LEDGF/p75, lens epithelium-derived growth factor; NT, non-targeting; PAC-Seq, Poly(A)-Click-Sequencing; UTR, untranslated region; WT, wild-type; hpi, hours post-infection.
    Figure Legend Snippet: Loss of CPSF6 expression globally shortens the 3’UTR of cellular mRNAs. ( A ) A549 WT, NT#H1, CPSF6-KO#B4, CPSF6-KO#B7, and CPSF6-KO#C8 cells were lysed, and proteins were analyzed by Nu-PAGE, followed by western blot using anti-CPSF6, anti-CPSF5, anti-CPSF7, and anti-GAPDH antibodies. Experiments were repeated at least three times, and a representative image is shown. Graphs show the average densitometry quantification of three replicates with standard deviation. Significance was determined using ANOVA multiple comparisons tests; *** P < 0.001; ns, not significant. ( B ) Total RNA from CPSF6-KO and WT A549 cells was prepared, and polyadenylated transcripts were identified by PAC-seq, followed by APA analysis using PolyAMiner. ( C ) A549 WT, NT#H1, CPSF6-KO#B4, CPSF6-KO#B7, and CPSF6-KO#C8 cells were infected with increasing amounts of HIV-1-GFP for 24 or 48 h. Infection was assessed as the percentage of GFP-positive cells by flow cytometry. ( D ) WT and CPSF6-KO#B7 A549 cells were infected with HIV-1-GFP at an MOI of 2 for 48 h. Cells were fixed, permeabilized, and stained using the following antibodies: (i) anti-CPSF5 (red) and anti-CPSF6 (green); (ii) anti-SC-35 (red) and anti-CPSF6 (green); (iii) anti-LEDGF/p75 (red) and anti-CPSF6 (green); and (iv) anti-LEDGF/p75 (red) and anti-SC35 (green). Nuclei were stained with DAPI (blue). Scale bar, 10 µm. ( E ) Percentage of A549 cells containing CPSF6, CPSF5, or CPSF7 in nuclear speckles (condensates) upon HIV-1 infection (average of three independent experiments with standard deviation). Cells containing CPSF6, CPSF5, or CPSF7 in nuclear speckles were determined by visual examination of 200 cells. Significance was determined using unpaired t -test; *** P < 0.001; ns, not significant. APA, alternative polyadenylation; CPSF5, cleavage and polyadenylation specificity factor subunit 5; CPSF6, cleavage and polyadenylation specificity factor subunit 6; CPSF7, cleavage and polyadenylation specificity factor subunit 7; DAPI, 4’,6-diamidino-2-phenylindole; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; KO, knockout; LEDGF/p75, lens epithelium-derived growth factor; NT, non-targeting; PAC-Seq, Poly(A)-Click-Sequencing; UTR, untranslated region; WT, wild-type; hpi, hours post-infection.

    Techniques Used: Expressing, Western Blot, Standard Deviation, Infection, Flow Cytometry, Staining, Knock-Out, Derivative Assay, Sequencing



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    Loss of CPSF6 expression globally shortens the 3’UTR of cellular mRNAs. ( A ) A549 WT, NT#H1, CPSF6-KO#B4, CPSF6-KO#B7, and CPSF6-KO#C8 cells were lysed, and proteins were analyzed by Nu-PAGE, followed by western blot using anti-CPSF6, anti-CPSF5, anti-CPSF7, and anti-GAPDH antibodies. Experiments were repeated at least three times, and a representative image is shown. Graphs show the average densitometry quantification of three replicates with standard deviation. Significance was determined using ANOVA multiple comparisons tests; *** P < 0.001; ns, not significant. ( B ) Total RNA from CPSF6-KO and WT A549 cells was prepared, and polyadenylated transcripts were identified by PAC-seq, followed by APA analysis using PolyAMiner. ( C ) A549 WT, NT#H1, CPSF6-KO#B4, CPSF6-KO#B7, and CPSF6-KO#C8 cells were infected with increasing amounts of HIV-1-GFP for 24 or 48 h. Infection was assessed as the percentage of GFP-positive cells by flow cytometry. ( D ) WT and CPSF6-KO#B7 A549 cells were infected with HIV-1-GFP at an MOI of 2 for 48 h. Cells were fixed, permeabilized, and stained using the following antibodies: (i) anti-CPSF5 (red) and anti-CPSF6 (green); (ii) anti-SC-35 (red) and anti-CPSF6 (green); (iii) anti-LEDGF/p75 (red) and anti-CPSF6 (green); and (iv) anti-LEDGF/p75 (red) and anti-SC35 (green). Nuclei were stained with DAPI (blue). Scale bar, 10 µm. ( E ) Percentage of A549 cells containing CPSF6, CPSF5, or CPSF7 in nuclear speckles (condensates) upon HIV-1 infection (average of three independent experiments with standard deviation). Cells containing CPSF6, CPSF5, or CPSF7 in nuclear speckles were determined by visual examination of 200 cells. Significance was determined using unpaired t -test; *** P < 0.001; ns, not significant. APA, alternative polyadenylation; CPSF5, cleavage and polyadenylation specificity factor subunit 5; CPSF6, cleavage and polyadenylation specificity factor subunit 6; CPSF7, cleavage and polyadenylation specificity factor subunit 7; DAPI, 4’,6-diamidino-2-phenylindole; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; KO, knockout; LEDGF/p75, lens epithelium-derived growth factor; NT, non-targeting; PAC-Seq, Poly(A)-Click-Sequencing; UTR, untranslated region; WT, wild-type; hpi, hours post-infection.

    Journal: mBio

    Article Title: HIV-1 infection regulates gene expression by altering alternative polyadenylation correlated with CPSF6 and CPSF5 redistribution

    doi: 10.1128/mbio.02865-25

    Figure Lengend Snippet: Loss of CPSF6 expression globally shortens the 3’UTR of cellular mRNAs. ( A ) A549 WT, NT#H1, CPSF6-KO#B4, CPSF6-KO#B7, and CPSF6-KO#C8 cells were lysed, and proteins were analyzed by Nu-PAGE, followed by western blot using anti-CPSF6, anti-CPSF5, anti-CPSF7, and anti-GAPDH antibodies. Experiments were repeated at least three times, and a representative image is shown. Graphs show the average densitometry quantification of three replicates with standard deviation. Significance was determined using ANOVA multiple comparisons tests; *** P < 0.001; ns, not significant. ( B ) Total RNA from CPSF6-KO and WT A549 cells was prepared, and polyadenylated transcripts were identified by PAC-seq, followed by APA analysis using PolyAMiner. ( C ) A549 WT, NT#H1, CPSF6-KO#B4, CPSF6-KO#B7, and CPSF6-KO#C8 cells were infected with increasing amounts of HIV-1-GFP for 24 or 48 h. Infection was assessed as the percentage of GFP-positive cells by flow cytometry. ( D ) WT and CPSF6-KO#B7 A549 cells were infected with HIV-1-GFP at an MOI of 2 for 48 h. Cells were fixed, permeabilized, and stained using the following antibodies: (i) anti-CPSF5 (red) and anti-CPSF6 (green); (ii) anti-SC-35 (red) and anti-CPSF6 (green); (iii) anti-LEDGF/p75 (red) and anti-CPSF6 (green); and (iv) anti-LEDGF/p75 (red) and anti-SC35 (green). Nuclei were stained with DAPI (blue). Scale bar, 10 µm. ( E ) Percentage of A549 cells containing CPSF6, CPSF5, or CPSF7 in nuclear speckles (condensates) upon HIV-1 infection (average of three independent experiments with standard deviation). Cells containing CPSF6, CPSF5, or CPSF7 in nuclear speckles were determined by visual examination of 200 cells. Significance was determined using unpaired t -test; *** P < 0.001; ns, not significant. APA, alternative polyadenylation; CPSF5, cleavage and polyadenylation specificity factor subunit 5; CPSF6, cleavage and polyadenylation specificity factor subunit 6; CPSF7, cleavage and polyadenylation specificity factor subunit 7; DAPI, 4’,6-diamidino-2-phenylindole; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; KO, knockout; LEDGF/p75, lens epithelium-derived growth factor; NT, non-targeting; PAC-Seq, Poly(A)-Click-Sequencing; UTR, untranslated region; WT, wild-type; hpi, hours post-infection.

    Article Snippet: We used rabbit polyclonal antibodies against the following proteins: CPSF6 (Cat# ab99347, Abcam), CPSF7 (Cat#A301-359A, Bethyl Laboratories, Inc), LEDGF/p75 (Cat# A300-847A, Bethyl Laboratories, Inc), SLFN5 (Cat#PA5-53638, Invitrogen), and MxA (Cat#PA5-22101, Invitrogen) and VMA21 (Cat#21921-1-AP, Proteintech).

    Techniques: Expressing, Western Blot, Standard Deviation, Infection, Flow Cytometry, Staining, Knock-Out, Derivative Assay, Sequencing