Fig. 3
From: CRISPR-Cas9-mediated labelling of the C-terminus of human laminin β1 leads to secretion inhibition
LMβ1::Dendra2 is expressed inside cells, but no secretion of LMβ1::Dendra2 was observed. a Total cell lysates were processed by western immunoblotting with anti-LMβ1 antibodies. Red box represents an enlarged section of the blot. Arrowed indicates additional upper LMβ1 band consistent with LMβ1::Dendra2. b Conditioned media and extracellular matrix lysates prepared for the indicated times then processed for immunoblotting with anti-LMβ1 antibodies. c, d Control A549 or 59B2 cells were seeded on coverslips for 48 h then either fixed and imaged with a z-stack (panel c) or processed for indirect immunofluorescence microscopy with antibodies against LMα5 (panel d). In e LMβ1::Dendra2 cells were cultured for 48 h on glass coverslips then removed with ammonium hydroxide to reveal only the extracellular matrix, then processed with antibodies against LMα5. f Cell cycle analysis of 59B2 cells (x’s) against wild type A549 cells (circles) analysed 24 h after serum shock. Proportion of cell population in each phase of the cell cycle was then plotted. Data was plotted in GraphPad. All scale bars represent 20 μm