Journal: The Journal of Clinical Investigation

Article Title: Targeting kinesin family member 20A sensitizes stem-like triple-negative breast cancer cells to standard chemotherapy

doi: 10.1172/JCI182394

Figure Lengend Snippet: ( A ) Top hits of KIF20A binding proteins identified by immunoprecipitation–mass spectrometry. ( B and C ) Co-IP of exogenous HA-tagged KIF20A and endogenous SMARCA4 in MDA-MB-231 ( B ) or HCC1806 ( C ) cells. ( D and E ) Immunoblot analysis of extracted nuclear and cytoplasmic proteins in MDA-MB-231 ( D ) or HCC1806 ( E ) cells. ( F ) Immunofluorescence staining of endogenous KIF20A and SMARCA4 in MDA-MB-231 or Hs587T cell lines. Scale bar: 90 μm.

Article Snippet: The cells were incubated with 1:1,000 dilution of KIF20A primary antibody (Proteintech, 15911-1-AP) or 1:800 dilution of SMARCA4 primary antibody (Proteintech, 66561-1-Ig) at 4°C overnight followed by 1:1,000 dilution of secondary antibody [Donkey anti-Rabbit IgG ReadyProbes, Alexa Fluor 488, Invitrogen, R37118 ; and Goat anti-Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 647, Invitrogen, A-21235].

Techniques: Binding Assay, Immunoprecipitation, Mass Spectrometry, Co-Immunoprecipitation Assay, Western Blot, Immunofluorescence, Staining

Journal: The Journal of Clinical Investigation

Article Title: Targeting kinesin family member 20A sensitizes stem-like triple-negative breast cancer cells to standard chemotherapy

doi: 10.1172/JCI182394

Figure Lengend Snippet: ( A – C ) Immunoblot analysis of SMARCA4 protein ( A ), RT-qPCR analysis of OXPHOS genes ( n = 3) ( B ), and measurement of OCR ( C ) in MDA-MB-231 cells transduced with indicated control or SMARCA4 shRNAs. ( D – F ) Immunoblot analysis of KIF20A and SMARCA4 ( D ) and RT-qPCR analysis of OXPHOS genes in MDA-MB-231 ( E ) and HCC1806 ( F ) cells transduced with control or KIF20A sgRNA and control or SMARCA4 shRNA as indicated. n = 3. ( G and H ) Measurement of OCR ( n = 5) ( G ) and mammosphere formation assay and corresponding quantifications ( n = 3) ( H ) in MDA-MB-231 cells transduced with control or KIF20A sgRNA and control or SMARCA4 shRNA as indicated. Scale bar: 1 mm. ( I ) ChIP-PCR analysis of KIF20A and SMARCA4 binding at the promoters of OXPHOS genes in MDA-MB-231 cells. n = 3. ( J ) ChIP-PCR analysis of SMARCA4 binding at the promoters of OXPHOS genes in MDA-MB-231 cells transduced with control or KIF20A sgRNAs. n = 3. ( K ) Schematic model of the mechanism proposed for this study. Each data point represents a biological replicate. Data represent mean ± SEM. Statistical analyses were conducted by 1-way ANOVA with Dunnett’s test ( B , I , and J ) or with Tukey’s test ( E , F , and H ). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: The cells were incubated with 1:1,000 dilution of KIF20A primary antibody (Proteintech, 15911-1-AP) or 1:800 dilution of SMARCA4 primary antibody (Proteintech, 66561-1-Ig) at 4°C overnight followed by 1:1,000 dilution of secondary antibody [Donkey anti-Rabbit IgG ReadyProbes, Alexa Fluor 488, Invitrogen, R37118 ; and Goat anti-Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 647, Invitrogen, A-21235].

Techniques: Western Blot, Quantitative RT-PCR, Transduction, Control, shRNA, Tube Formation Assay, Binding Assay

Journal: iScience

Article Title: BRG1 is involved in vascular calcification in chronic renal disease via autophagy of vascular smooth muscle cells

doi: 10.1016/j.isci.2023.106485

Figure Lengend Snippet: Key resources table

Article Snippet: Enzyme linked immunosorbent assay (ELISA) kits (BRG1) , CUSABIO , Cat#CSB-EL021801HU.

Techniques: Recombinant, Modification, Saline, Lysis, Marker, Enzyme-linked Immunosorbent Assay, Immunohistochemistry, Preserving, Plasmid Preparation, Software, Imaging