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. Author manuscript; available in PMC: 2019 Sep 1.
Published in final edited form as: J Neurochem. 2018 Aug 2;146(5):526–539. doi: 10.1111/jnc.14463

Figure 9. Knockdown of GPR75 decreases CCL5-induced pERK.

Figure 9.

A. SH-SY5Y cells were transfected with CRISPR/Cas9 components once per cell passage (4 cell passages total) to enrich GPR75 knockdown within the population. After confirmation of GPR75 knockdown, cells were split and plated onto a 12 well plate. After 24 hours cells were stimulated with CCL5 (50 ng/ml) for the specified time-points. Cell lysates were separated on a gel for western blot analysis of pERK1/2 and total ERK1/2. B. Densitometric analysis of the ratio of pERK1/2-total ERK1/2. Data are the mean + SD of three independent samples for each time-point. *p<0.05 GPR75 gRNA A vs. WT, #p<0.05 GPR75 gRNA B vs. WT. C. An example of a western blot stripped and re-probed for GPR75 and β-Actin to confirm protein knockdown. D. Densitometric analysis shows significant decrease of GPR75 expression in mutant cell lines. *p<0.05 GPR75 gRNA A vs. WT, #p<0.05 GPR75 gRNA B vs. WT. Data, expressed in AU, are the mean + SD of three independent samples for each treatment.