Key resources table.
| Reagent type (species) or resource |
Designation | Source or reference |
Identifiers | Additional information |
|---|---|---|---|---|
| Gene (Mus musculus) | Krt6a | UniProtKB - P50446 | Targeted by CRISPR/Cas9 Knockout | |
| Gene (Mus musculus) | Krt6b | UniProtKB - Q9Z331 | Targeted by CRISPR/Cas9 Knockout | |
| Strain, strain background (E. coli) | DH5α | ThermoFisher Scientific | Cat# 18265017 | Chemically competent cells |
| Cell line (Mus musculus) | KtyI KO K14 | Homberg et al., 2015. DOI: 10.1038/jid.2015.184 | T. Magin lab, Institute of Biology, University of Leipzig, Germany. Cell line used to generate K5/K14_1 cell line |
|
| Cell line (Mus musculus) | K5/K14_1 | This paper | Clonal knockout cell line of Krt6a and Krt6b, maintained in the Medalia lab | |
| Antibody | Anti-mouse Keratin 14 (mouse monoclonal) | ThermoFisher Scientific | MA5-11599, Clone LL002 | (1:100 – 1:10) |
| Antibody | Anti-mouse Keratin 5 (rabbit polyclonal) | BioLegend | Cat# 905503 | (1:500) |
| Antibody | Anti-mouse Keratin 6a (rabbit polyclonal) | BioLegend | Cat# 905702 | (1:500) |
| Antibody | Cy3 AffiniPure anti-rabbit (donkey polyclonal) | Jackson Immuno Research | Cat# 711-165-152 | (1:100) |
| Antibody | FITC AffiniPure anti-mouse (donkey polyclonal) | Jackson Immuno Research | Cat# 715-095-150 | (1:100) |
| Recombinant DNA reagent | pX458 (pSpCas9(BB)−2A-GFP) (plasmid) | Addgene | Cat# 48138 | CRISPR/Cas9 knockout |
| Recombinant DNA reagent | pGEM-T Easy (plasmid) | Promega | Cat# A1360 | |
| Sequence-based reagent | guideRNA insert targetting Krt6a and Krt6b gene | This paper | guideRNA sequence: GAGCCACCGCTGCCCCGGGAG. guideRNA cloned into pX458 plasmid and transfected into KtyI KO K14 cells. | |
| Commercial assay or kit | P3 primary cell 4D-Nucleofector kit | Lonza | Cat# V4XP-3032 | Transfection of KtyI KO K14 cells |
| Commercial assay or kit | jetPRIME | Polyplus transfection | Cat# 114–07 | Transfection of KtyI KO K14 cells |
| Commercial assay or kit | GenElute Mammalian Genomic DNA kit | Sigma-Aldrich | Cat# G1N70-1KT | Genomic DNA extraction |
| Chemical compound, drug | 99.9% anhydrous methanol | Alfa Aesar | Cat# 41838 | Fixation of mammalian cells |
| Chemical compound, drug | Hoechst 33342 | Sigma-Aldrich | Cat# B2261 | (1:10000) |
| Chemical compound, drug | Dako mounting medium | Agilent | Cat# S3023 | |
| Chemical compound, drug | Prolong Glass Anti-Fade | ThermoFisher Scientific | Cat# P36980 | |
| Software, algorithm | TIDE webtool | Netherlands Cancer Institute | https://tide.nki.nl/ | CRISPR/Cas9 knockout analysis |
| Software, algorithm | FIJI | Max Planck Institute of Molecular Cell Biology and Genetics, Dresden, Germany | RRID:SCR_002285 | For data analysis |
| Software, algorithm | Illustrator | Adobe Inc | RRID:SCR_010279 | For data analysis |
| Software, algorithm | OriginPro | OriginLab | RRID:SCR_014212 | For data analysis |
| Software, algorithm | MATLAB | MathWorks | RRID:SCR_001622 | For data analysis |
| Software, algorithm | RELION | Scheres, 2012. DOI: 10.1016/j.jsb.2012.09.006 | RRID:SCR_016274 | For data analysis |
| Software, algorithm | IMOD | University of Colorado Boulder, Colorado, USA | RRID:SCR_003297 | For data analysis |
| Software, algorithm | Amira | ThermoFisher Scientific | RRID:SCR_007353 | For data analysis |
| Software, algorithm | SerialEM | University of Colorado Boulder, Colorado, USA | RRID:SCR_017293 | For data acquisition |
| Software, algorithm | UCSF Chimera | University of California, California, USA | RRID:SCR_004097 | For structure visualization |