rabbit α cnot1  (Cell Signaling Technology Inc)

Journal: International Journal of Molecular Sciences

Article Title: Stress-Induced Changes in Nucleocytoplasmic Localization of Crucial Factors in Gene Expression Regulation

doi: 10.3390/ijms25073895

Figure Lengend Snippet: Cytoplasmic localization ( A ) and expression level ( B ) of CNOT1 among control (n Ctr = 8) and smoking origin derived (n Sm = 6) umbilical cord vessels. Dot plots showed the semiquantified results of CNOT1 specific immunolabeling on the arterial and venous endothelial cells. Dots and rhombuses ( A ) symbolize the overall MFI values in percentile for the cytoplasmic distribution of individual samples. Fold of changes in the total MFI values ( B ), where the summarized data from control group were applied as the baseline of comparison (mean ± SEM). Statistics: unpaired t -test (two-tailed) followed by Mann-Whitney test to compare ranks (* p ≤ 0.05, **** p ≤ 0.0001).

Article Snippet: Primary antibodies: rabbit anti-PARP-1 monoclonal (C.384.8): MA5-15031; Invitrogen (Waltham, MA, USA), mouse anti-cleaved PARP-1 monoclonal (194C1439): sc-56196; Santa Cruz Biotechnology (Dallas, TX, USA), rabbit anti-CNOT1 polyclonal: ab234642; Abcam (Cambridge, UK), mouse anti-Pol II (RPB1) monoclonal (A-10): sc-17798 Santa Cruz Biotechnology (Dallas, TX, USA), rabbit, anti-cleaved Casp3 polyclonal: ab2302; Abcam (Cambridge, UK).

Techniques: Expressing, Derivative Assay, Immunolabeling, Comparison, Two Tailed Test, MANN-WHITNEY

Journal: International Journal of Molecular Sciences

Article Title: Stress-Induced Changes in Nucleocytoplasmic Localization of Crucial Factors in Gene Expression Regulation

doi: 10.3390/ijms25073895

Figure Lengend Snippet: Stress induces changes in the distribution of CNOT1 within the cytoplasm, which impacts the assembly of RNA polymerase II and its subsequent transport into the nucleus, leading to increased levels of PARP-1 (created with Biorender.com). ( A ): Under normal circumstances, CNOT1 typically exhibits a granular pattern within the cytoplasm. The assembly of Rpb1 and Rpb2, crucial for their translocation into the nucleus, depends on CNOT1. However, the granular association of CNOT1, likely associated with assemblysomes, results in limited accumulation of Rpb1 in the nucleus. Consequently, transcription and subsequent expression of proteins such as PARP-1 remain low. ( B ): When exposed to stress, CNOT1 demonstrates a more dispersed cytoplasmic staining, suggesting a shift from assemblysomes to polysomes. This transition increases the availability of soluble CNOT1 for the assembly of Rpb1 and Rpb2, facilitating their nuclear accumulation. As a result, the relocation of RPB1 to the nucleus enhances overall transcription, leading to elevated levels of PARP-1 proteins.

Article Snippet: Primary antibodies: rabbit anti-PARP-1 monoclonal (C.384.8): MA5-15031; Invitrogen (Waltham, MA, USA), mouse anti-cleaved PARP-1 monoclonal (194C1439): sc-56196; Santa Cruz Biotechnology (Dallas, TX, USA), rabbit anti-CNOT1 polyclonal: ab234642; Abcam (Cambridge, UK), mouse anti-Pol II (RPB1) monoclonal (A-10): sc-17798 Santa Cruz Biotechnology (Dallas, TX, USA), rabbit, anti-cleaved Casp3 polyclonal: ab2302; Abcam (Cambridge, UK).

Techniques: Translocation Assay, Expressing, Staining