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. Author manuscript; available in PMC: 2019 Jun 9.
Published in final edited form as: Cancer Res. 2018 Mar 13;78(10):2490–2502. doi: 10.1158/0008-5472.CAN-17-2392

Figure 2. Knockout of CaMKII in PC3-mm2 cells.

Figure 2.

PC3-mm2 cells were transfected with empty vector (vector) or CRISPR/Cas9 targeting CaMKII genes (clones #1, #6, #10). (A) Total cell lysates were immunoblotted for pCaMKII and total CaMKII. GAPDH was used as a loading control. (B) Cells were cultured in 1% FBS and cell proliferation was measured by cell counting. The results were expressed as fold change of cell number counted on Day 0. n=3. The doubling time of the vector control PC3-mm2 cells was 18.2 ± 1.2 hours, and those for the CaMKII knockout clones #1, #6, #10 were 32.1 ± 1.0 hours, 43.5 ± 11.9 hours, and 35.6 ± 0.2 hours, respectively. The plating efficiency, determined at 24 hours after cell seeding, of the vector control cells was 87.5%, and for the CaMKII knockout clones #1, #6 and #10 were 70.7%, 86.5% and 76.0%, respectively. (C) Cells were grown in soft agar. Number of colonies per field was counted. n=3. (D) Cells were seeded onto a Boyden chamber. Cells migrated through the membrane were labeled with Calcein AM. n=2. (E) Cells were seeded into BioCoat Matrigel-coated invasion chamber. Cells invaded through the matrigel were labeled with Calcein AM. n=3. *, p<0.05, **, p<0.01, ***, p<0.001.