Journal: Cell stem cell

Article Title: A CLK3-HMGA2 alternative splicing axis impacts human hematopoietic stem cell molecular identity throughout development

doi: 10.1016/j.stem.2018.03.012

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: DGCR8 knockout MEF , Novus Biologicals , Cat. NBP2-25171.

Techniques: Recombinant, Transfection, Reporter Assay, DNA Library Preparation, Expressing, Knock-Out, Negative Control, Software

Journal: Cell Death & Disease

Article Title: Protein disulfide isomerase blocks the interaction of LC3II-PHB2 and promotes mTOR signaling to regulate autophagy and radio/chemo-sensitivity

doi: 10.1038/s41419-022-05302-w

Figure Lengend Snippet: A – D Immunoblot analysis showing the levels of ERS pathway-related proteins GRP78, PERK, ATF6 ( A , C ) or AKT/mTOR pathway-related proteins ( B , D ) in PDI knockdown or overexpression HCT116 or MEF cells treated with γ-ray (4 Gy) or cisplatin (20 μM) irradiation for 24 h. E , F Immunoblot analysis showing the levels of ERS pathway-related proteins and AKT/mTOR pathway-related proteins in PDI knockdown or overexpression MEF cells treated with γ-ray irradiation ( E ) or cisplatin ( F ) for 24 h. G , H The existence of PDI in the co-precipitated complexes was confirmed by western blotting, IgG was employed as the negative control. Western blotting shows the association of PDI with GRP78 after Co-IP in HCT116 cells were treated with γ-ray (4 Gy) irradiation ( G ) or cisplatin (20 μM) ( H ) for 24 h. I , J The existence of PDI in the co-precipitated complexes was confirmed by western blotting, IgG was employed as the negative control. Western blotting shows the association of PDI with GRP78 after Co-IP in MEF cells were treated with γ-ray (4 Gy) irradiation ( I ) or cisplatin (20 μM) ( J ) for 24 h. K , L HCT116 cells were pretreated with or without 5 mM MK2206 ( K ) or 10 µM 4-PBA ( L ) for 2 h, then irradiated with 4 Gy γ-ray for 24 h. LC3II accumulation was measured using western blotting.

Article Snippet: The human cell lines HCT116, HEK293T, A549, HIEC, and MEF cells were kindly provided by Dr Chi Li (University of Louisville, Kentucky, USA).

Techniques: Western Blot, Knockdown, Over Expression, Irradiation, Negative Control, Co-Immunoprecipitation Assay

Journal: Cell Death & Disease

Article Title: Protein disulfide isomerase blocks the interaction of LC3II-PHB2 and promotes mTOR signaling to regulate autophagy and radio/chemo-sensitivity

doi: 10.1038/s41419-022-05302-w

Figure Lengend Snippet: A – D The interaction between PDI and PHB2 was detected by immunoprecipitation after the treatment with γ-ray (4 Gy) irradiation ( A , B ) or 20 μM cisplatin ( C , D ) for 24 h in HCT116 and MEF cells. E , F Immunofluorescence microscope analysis of co-localization of PDI (red) and PHB2 (green) in PDI knockdown or PDI overexpression HCT116 cells after 24 h of treatment with IR (4 Gy) ( E ) or cisplatin (20 μM) ( F ). Blue DAPI staining was used to stain the cell nucleus. Scale bar = 25 μm. G The interaction between PDI and PHB2 was detected by Flag-pull down in vitro. Purified Flag-PHB2 or Myc-PDI immobilized on the beads was incubated with purified Myc-PDI or Flag-PHB2. Input and bead-bound proteins were analyzed by immunoblotting with anti-PDI or anti-PHB2 antibodies. H The changes in the MMP of HCT116 cells before and after γ-ray (4 Gy) irradiation for 24 h were detected by the JC-1 probe. I Statistical map of MMP. J The expression of ROS in HCT116 cells after γ-ray (4 Gy) irradiation for 24 h. K , L Expression of mitophagy-related proteins in mitochondria treated with γ-ray (4 Gy) irradiation ( K ) after 24 h in HCT116 cells or treated with cisplatin (20 μM) after 24 h in MEF cells ( L ). Data were pooled from three independent experiments and the results are represented as mean ± SD; * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The human cell lines HCT116, HEK293T, A549, HIEC, and MEF cells were kindly provided by Dr Chi Li (University of Louisville, Kentucky, USA).

Techniques: Immunoprecipitation, Irradiation, Immunofluorescence, Microscopy, Knockdown, Over Expression, Staining, In Vitro, Purification, Incubation, Western Blot, Expressing