Journal: Cell Host & Microbe

Article Title: Determinants of Spike infectivity, processing, and neutralization in SARS-CoV-2 Omicron subvariants BA.1 and BA.2

doi: 10.1016/j.chom.2022.07.006

Figure Lengend Snippet:

Article Snippet: Thereafter, samples were washed with PBS and incubated for 2 h at 4°C with primary antibody (anti-V5(Mouse) (1:1,000, Cell Signalling, #80076S)) diluted in PBS.

Techniques: Recombinant, Mutagenesis, Binding Assay, Plasmid Preparation, Software, Microscopy, Modification

Journal: bioRxiv

Article Title: Condensin chromatin association is regulated by SMC head and hinge engagement and phosphorylation

doi: 10.1101/2025.11.17.688780

Figure Lengend Snippet: Expression of inducible Smc2 mutants created by conditional protein exchange. A) Schematic view of budding yeast condensin, illustrating the relative locations of each Smc2 mutation. Engagement of ATP-bound head domains triggers ATP hydrolysis, leading to their disengagement. The K38I and D1112A mutations are predicted to block binding of ATP to their respective pockets. The E113Q mutation is predicted to slow down the rate of ATP hydrolysis, while R58A is predicted to disrupt DNA dependent ATP hydrolysis. The S1085R mutation is predicted to disrupt head-head interactions. B) Within the hinge region of Smc2, we designed two mutations to specifically disrupt either of the two interfaces made with Smc4: hinge interface 1, L665R; hinge interface 2, L567K. Inset panels show additional details for how each single point mutant is predicted to generate a series of steric clashes (red circles) incompatible with hetero-dimerization of the hinge. Key amino acids of Smc2 and Smc4 are labelled in plain and italic text respectively. Figures were generated using MacPyMOL. C) Conditional protein exchange system for smc2 mutations. Under the permissive conditions (2% raffinose, 25°C) functional degron and HA tagged protein is expressed. Under the restrictive conditions (+2% galactose, + doxycycline 37°C), the functional degron protein is degraded and the exogenous HA tagged protein is expressed from a GAL1 promoter. D) Spot test of viability following conditional protein exchange for the different smc2 mutants. E) Condensin complexes generated following expression of different smc2 mutants. Following conditional protein exchange, in vivo condensin complexes were immuno-precipitated and the isolated complexes analyzed by MS/MS as described in the Methods. The signal from several peptides derived from each protein was quantitated by MS/MS and then normalized to the quantity of peptides from the Brn1 subunit. This provides a measure of the relative quantity of each subunit immuno-precipitated by Brn1-V5. All experiments are the average of two independent experiments and the standard deviation of the experiment shown by the error bar.

Article Snippet: Anti-V5 antibody (mouse monoclonal MCA1360, abD Serotec) used at 1:1000 Concentration in Western.

Techniques: Expressing, Mutagenesis, Blocking Assay, Binding Assay, Generated, Functional Assay, Spot Test, In Vivo, Isolation, Tandem Mass Spectroscopy, Derivative Assay, Standard Deviation

Journal: bioRxiv

Article Title: Condensin chromatin association is regulated by SMC head and hinge engagement and phosphorylation

doi: 10.1101/2025.11.17.688780

Figure Lengend Snippet: Chromatin association of condensin following expression of smc2 enzymatic mutations. Conditional expression depletion strains were arrested in G1 and endogenous Smc2 degraded and the indicated version of exogenous Smc2 expressed. Cells were then released from G1 into restrictive medium containing nocodazole. After 2 h, cells were harvested, and Brn1-V5 levels at the indicated sites analyzed by ChIP-qPCR. The average of at least three experimental repeats (qPCR performed in triplicate in each case) is shown with error bars. Enrichments are grouped according to function,

Article Snippet: Anti-V5 antibody (mouse monoclonal MCA1360, abD Serotec) used at 1:1000 Concentration in Western.

Techniques: Expressing, ChIP-qPCR

Journal: bioRxiv

Article Title: Condensin chromatin association is regulated by SMC head and hinge engagement and phosphorylation

doi: 10.1101/2025.11.17.688780

Figure Lengend Snippet: Effects of phosphorylation defective condensin subunits on chromatin association. Strains expressing the defined phosphorylation mutants of A) Smc4- smc4-10A , B) Brn1- brn1-570 , C) Ycs4 – ycs4-543 D) Ycg1, ycg1-521 were released from G1 into medium containing nocodazole at 37°C. After 2 h, cells were harvested, and Brn1-V5 levels at the indicated sites analyzed by ChIP-qPCR. The average of at least three experimental repeats (qPCR performed in triplicate in each case) is shown with error bars.

Article Snippet: Anti-V5 antibody (mouse monoclonal MCA1360, abD Serotec) used at 1:1000 Concentration in Western.

Techniques: Phospho-proteomics, Expressing, ChIP-qPCR

Journal: bioRxiv

Article Title: Condensin chromatin association is regulated by SMC head and hinge engagement and phosphorylation

doi: 10.1101/2025.11.17.688780

Figure Lengend Snippet: Requirement for Ipl1 and Cdc5 for condensin chromatin association and complex stability. Either A) ipl1-321 , B) cdc5-10 C) cdc5-99 cells were released from G1 into medium containing nocodazole at 37°C. After 2 h, cells were harvested, and Brn1-V5 levels at the indicated sites analyzed by ChIP-qPCR. The average of at least three experimental repeats (qPCR performed in triplicate in each case) is shown with error bars.

Article Snippet: Anti-V5 antibody (mouse monoclonal MCA1360, abD Serotec) used at 1:1000 Concentration in Western.

Techniques: ChIP-qPCR

Journal: The Journal of Biological Chemistry

Article Title: Secreted Hsp90 Is a Novel Regulator of the Epithelial to Mesenchymal Transition (EMT) in Prostate Cancer *

doi: 10.1074/jbc.M112.389015

Figure Lengend Snippet: Modest elevation of eHsp90 is sufficient to suppress E-cadherin function and promote cell motility. A, upper panel, ELISA analysis of secreted eHsp90 protein detected from conditioned media collected from parental ARCaPE cells stably transduced with control (lacZ) or V5-tagged eHsp90α lentivirus. Lower panel, immunoblot detection of total (endogenous and exogenous) eHsp90α, or V5 detection of transduced eHsp90 protein. B, immunoblot analysis of cell lysates from ARCaPE-LacZ or ARCaPE-eHsp90 confirmed consistent levels of intracellular eHsp90α (IC Hsp90). Indicated analysis of E- and N-cadherin and ERK activity. C, representative morphology of the indicated ARCaPE cells. Analysis of cell motility of ARCaPE-eHsp90 either untreated or treated with NPGA. D, effect of NPGA upon E-cadherin expression in ARCaPE-eHsp90. E, analysis of E-cadherin localization in ARCaPE-LacZ and ARCaPE-eHsp90 untreated cells, or treated for the indicated times with NPGA. F, membrane localization of ZO1 in ARCaPE-LacZ and ARCaPE-eHsp90 untreated cells, or treated with NPGA for 3 days. Asterisks (*) indicate significance of p value ≤0.05. Scale bar is 50 μm.

Article Snippet: V5 antibody (NB600-381) was from Novus Biologicals.

Techniques: Enzyme-linked Immunosorbent Assay, Stable Transfection, Transduction, Western Blot, Activity Assay, Expressing