mouse anti cpsf3  (Santa Cruz Biotechnology)

Journal: Cell reports

Article Title: Non-canonical isoforms of the mRNA polyadenylation factor WDR33 regulate STING-mediated immune responses

doi: 10.1016/j.celrep.2024.113886

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Mouse anti-CPSF3 , Santa Cruz Biotechnology , Cat# sc-393001.

Techniques: Virus, Recombinant, Negative Control, Software

Journal: iScience

Article Title: Targeting the mRNA endonuclease CPSF73 inhibits breast cancer cell migration, invasion, and self-renewal

doi: 10.1016/j.isci.2022.104804

Figure Lengend Snippet: UBE3D positively regulates CPSF73 but not other C/P proteins at the post-translational level (A) Western blot verification at the protein level of UBE3D KO in HEK293 cells and UBE3D addback to KO cells. (B) CPSF73 was depleted in UBE3D KO cells and its level was restored by adding back UBE3D. Other C/P proteins were not depleted. A representative Western blot performed using antibodies specific to the subunits of the C/P complex is shown on the left panel. Quantitation of the log2-fold changes in protein levels compared to wild-type (WT) using two replicates is shown on the right. (C) CPSF73 mRNA level remained unchanged in UBE3D -negative and positive cells. Three biological replicates were analyzed by RT-qPCR, and values normalized to RPL13A expression. (D) Cycloheximide (CHX) chase assay revealed that CPSF73 is less stable in UBE3D KO cells. Left panel, representative Western Blot and right panel, quantification of protein levels from three replicates, with values normalized to GAPDH. (E) The level of CPFS73 in UBE3D KO cells was increased by the proteasome inhibitor MG132 (10 μM) and the E1 ubiquitin-activating enzyme inhibitor TAK243 (1 μM), demonstrating the degradation of CPFS73 was mediated by the ubiquitin-proteasome pathway. CPSF73 protein level with each treatment was normalized to the untreated control from two replicates. For all panels, error bars show the mean ± SEM; ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ∗∗∗∗p < 0.0001 (Student’s t test, unpaired, two-tailed).

Article Snippet: Mouse monoclonal anti-CPSF73 antibody (C-3) , Santa Cruz Biotechnology , Cat#sc-393001.

Techniques: Western Blot, Quantitation Assay, Quantitative RT-PCR, Expressing, Ubiquitin Proteomics, Control, Two Tailed Test

Journal: iScience

Article Title: Targeting the mRNA endonuclease CPSF73 inhibits breast cancer cell migration, invasion, and self-renewal

doi: 10.1016/j.isci.2022.104804

Figure Lengend Snippet: UBE3D depletion inhibits breast cancer cell motility in a CPFS73-dependent manner (A) Analysis of mRNA 3′ end processing efficiency of ACTB and MYC transcripts in MDA-MB-231 after UBE3D KD from two biological replicates, as described in Figure 2 A. (B) GO term analysis of the genes downregulated in MDA-MB-231 UBE3D KD samples was performed using Metascape. (C) Migration and invasion assessments in MDA-MB-231 cells upon UBE3D KD. Representative images of the migrated and invaded cells. Scale bar = 200 μm. (D) Quantification of migratory and invasive cells upon UBE3D KD. (E) CPSF73 overexpression in UBE3D KD cells was determined by Western blot. (F and G) CPSF73 overexpression reversed the UBE3D KD-mediated MDA-MB-231 migration and invasion defects. Scale bar = 200 μm. For all panels, error bars show the mean ± SEM; ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ∗∗∗∗p < 0.0001 (Student’s t test, unpaired, two-tailed).

Article Snippet: Mouse monoclonal anti-CPSF73 antibody (C-3) , Santa Cruz Biotechnology , Cat#sc-393001.

Techniques: Migration, Over Expression, Western Blot, Two Tailed Test

Journal: iScience

Article Title: Targeting the mRNA endonuclease CPSF73 inhibits breast cancer cell migration, invasion, and self-renewal

doi: 10.1016/j.isci.2022.104804

Figure Lengend Snippet: Catalytic inhibition of CPSF73 leads to breast cancer cell migration and invasion defects (A) ACTB and MYC mRNA 3′ end processing in MDA-MB-231 cells treated with JTE-607, determined from three replicates. (B and C) Transwell assays to evaluate the effects of JTE-607 treatment on cell migration and invasion. Representative images of the migrated and invaded cells (B) and quantitation data (C). Scale bar = 200 μm. (D) JTE-607 reduced MDA-MB-231 cell branches in a 3D invasion assay. Right panel: quantification of branches from 30 to 50 colonies in each group. (E) JTE-607 effectively inhibited MDA-MB-231 sphere formation in Matrigel. Spheres with a diameter over 50 μm were counted and quantified from three replicates. Representative images are shown. For all panels, error bars show the mean ± SEM; ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ∗∗∗∗p < 0.0001 (Student’s t test, unpaired, two-tailed).

Article Snippet: Mouse monoclonal anti-CPSF73 antibody (C-3) , Santa Cruz Biotechnology , Cat#sc-393001.

Techniques: Inhibition, Migration, Quantitation Assay, Invasion Assay, Two Tailed Test

Journal: iScience

Article Title: Targeting the mRNA endonuclease CPSF73 inhibits breast cancer cell migration, invasion, and self-renewal

doi: 10.1016/j.isci.2022.104804

Figure Lengend Snippet: CPSF73 expression level correlates with breast cancer cell self-renewal properties (A) CPSF73 knockdown by three independent inducible shRNAs in MDA-MB-231 cells. (B and C) CPSF73 knockdown effectively inhibited MDA-MB-231 sphere formation in Matrigel. Spheres with a diameter over 50 μm were counted and quantified from three replicates. Representative images are shown. Scale bar = 200 μm. (D) Doxycycline-inducible overexpression of CPSF73 in MDA-MB-231 cells. (E and F) CPSF73 overexpression (OE) promoted MDA-MB-231 sphere formation in Matrigel. Spheres with a diameter over 50 μm were counted and quantified from three replicates. Representative images are shown. Scale bar = 200 μm. For all panels, error bars show the mean ± SEM; ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ∗∗∗∗p < 0.0001 (Student’s t test, unpaired, two-tailed).

Article Snippet: Mouse monoclonal anti-CPSF73 antibody (C-3) , Santa Cruz Biotechnology , Cat#sc-393001.

Techniques: Expressing, Knockdown, Over Expression, Two Tailed Test

Journal: iScience

Article Title: Targeting the mRNA endonuclease CPSF73 inhibits breast cancer cell migration, invasion, and self-renewal

doi: 10.1016/j.isci.2022.104804

Figure Lengend Snippet:

Article Snippet: Mouse monoclonal anti-CPSF73 antibody (C-3) , Santa Cruz Biotechnology , Cat#sc-393001.

Techniques: Recombinant, Cell Viability Assay, shRNA, Control, Plasmid Preparation, Software