Fig. 2: Maspin distributes along cytoskeleton-enriched structures in MCF-10A cells.

a Maspin detected at the cell cortex during cell–cell contact re-establishment. Confluent GFP-Puro-N1-Maspin knock-in cells (left panel) were treated with 4 mM EGTA for 45 min, followed by recovery in complete medium for 1 h, fixed and co-stained for F-actin and α-tubulin. WT versus KO cells were treated identically and co-stained for Maspin, F-actin and α-tubulin (middle and right panels); (b) Maspin detected at the lamellipodia. Clustered GFP-Puro-N1-Maspin knock-in cells exhibiting discreet lamellipodial protrusions were fixed and co-stained for F-actin and α-tubulin (left panel). Clustered WT and KO cells were co-stained for Maspin, F-actin and α-tubulin (middle and right panels); (c) Maspin detected at the mitotic spindle. Asynchronous mitotic GFP-Puro-N1-Maspin knock-in cells were fixed and co-stained for F-actin and α-tubulin (left panel). WT versus KO cells were co-stained for Maspin, F-actin and α-tubulin (middle and right panels). Nuclei were stained with Hoechst. Individual Z-slices are depicted in the figure. Scale bars: 10 µm.