Journal: Autophagy

Article Title: Regulation of N-degron recognin-mediated autophagy by the SARS-CoV-2 PLpro ubiquitin deconjugase

doi: 10.1080/15548627.2024.2442849

Figure Lengend Snippet: Reagents used in the investigation.

Article Snippet: pEGFP N1 Ub-CRT , Addgene , 69619; Marta Hallak lab.

Techniques: Recombinant, Diagnostic Assay, DC Protein Assay, Transfection, Mutagenesis, Plasmid Preparation, Gel Extraction, Purification, Cell Culture, shRNA

Journal: Nature communications

Article Title: Multiple structures of RNA polymerase II isolated from human nuclei by ChIP-CryoEM analysis.

doi: 10.1038/s41467-025-59580-x

Figure Lengend Snippet: Fig. 2 | Details of the EC-SPT4/5-ELOF1-SPT6 structure. a Domain architectures of the elongation factors. The known domains are indicated. b Close-up view of ELOF1 (purple). c Close-up view around the upstream DNA. d Close-up view of the SPT5 KOW2, KOW3, KOWx, and KOW4 domains. Each KOW domain is indicated with dotted circles. SPT6 is omitted for clarity. e Close-up view of the KOW5 domain. f The SPT6-SPT5 KOW3-RPB4/7 stalk interaction. g The SPT5 KOW1-SPT6

Article Snippet: Themembraneswerewashedwith TBS-T (20mMTris-HCl (pH 7.5), 137mM NaCl, and 0.1% Tween-20) three times and then incubated overnight with anti-FLAG M2 antibody (Sigma # F3165, 1:3000), anti-SPT5 antibody (Proteintech#16511-1-AP, 1:1000), antiSPT6 antibody (Cell Signaling #15616, 1:2000), anti-PAF1 antibody (Cell Signaling #12883, 1:300), anti-RTF1 antibody (Proteintech #12170-1-AP, 1:1000), anti-MED17 antibody (Proteintech#11505-1-AP, 1:2000), or anti-XPB antibody (Cell Signaling #8746, 1:300) in Can Get Signal Immunoreaction Enhancer Solution 1 (TOYOBO #NKB-101).

Techniques:

Journal: Nature communications

Article Title: An unexpected protein interaction promotes drug resistance in leukemia.

doi: 10.1038/s41467-017-01678-y

Figure Lengend Snippet: Fig. 3 Peptides that block the ABCC4–MPP1 protein interaction reduce ABCC4 membrane localization and MPP1-driven hematopoietic progenitor replating in methylcellulose. a Representative image of ABCC4 membrane localization in cells transfected with ABCC4-GFP and MPP1 treated with either a cell penetrating peptide harboring the ABCC4-PDZ motif or lacking the PDZ motif (ABCC4ΔC). Blue-wheat germ agglutinin conjugated with Alexa Fluor 647, Red = Calreticuin-RFP, and Green = ABCC4-GFP. b Quantification of ABCC4 localization (n = 20 fields of view; bars represent SEM). c HPC were transduced with retroviral plasmid containing cDNAs for MPP1-ires-CFP or the empty vector, purified by FAC sorting and plated in methycellulose medium containing IL3, IL6, SCF, EPO and supplemented with cell penetrating peptide harboring the ABCC4-PDZ motif or lacking the PDZ-motif (ABCC4ΔC). Colonies were counted after 7 days growth and serially replated. Fresh peptide was added at each replating step (n = 3, bars represent SEM)

Article Snippet: HEK293 cells were seeded on chambered coverglass slides (LAB-TEK) 48 h before transfection with plasmid DNA (250 ng) encoding pAc-GFP-ABCC4 or ABCC4ΔC and pCal plasmid encoding the ER resident protein calreticulin conjugated to RFP (Origene).

Techniques: Blocking Assay, Membrane, Transfection, Transduction, Retroviral, Plasmid Preparation