Figure 6.

Detection of Laminin Subunit Alpha 1 Protein by Immunocytochemistry in C2C12 Cells

C2C12 cells were transfected with pAC154 coding for dCas9-VP160-2A-puro and a gRNA. The cells were selected with puromycin, expanded during 7 days and fixed with ethanol 95%. (A–F) The laminin subunit alpha 1 protein was detected by immunofluorescence (red label). The nuclei were stained with Hoechst 33258. (E and F) The negative control cells were transfected with a plasmid pAC154 not coding for a gRNA. (G) Proteins were extracted 18 days later, and laminin subunit alpha 1 protein was detected by western blot. (H) The best induction of the laminin subunit alpha 1 protein was observed in the cells transfected with gRNA3 and gRNA4. No laminin subunit alpha 1 protein was detected in the negative control cells transfected either with pAC154 not coding for a gRNA or a plasmid coding for eGFP.