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. Author manuscript; available in PMC: 2023 Jan 3.
Published in final edited form as: Sci Signal. 2022 Sep 27;15(753):eabk1147. doi: 10.1126/scisignal.abk1147

Figure 4. SCA14 mutants are resistant to phorbol ester-mediated downregulation.

Figure 4.

(A) Western blot of Triton-soluble lysates from COS7 transfected with HA-tagged WT PKCγ, PKCγ lacking a C1A domain (ΔC1A), PKCγ lacking a C1B domain (ΔC1B), the indicated SCA14 mutants, or with empty vector (Mock). Membranes were probed with anti-HA (PKCγ) or phospho- specific antibodies. N = three independent experiments. (B) Western blot of whole-cell lysates from COS7 cells transfected with HA-tagged WT PKCγ, PKCγ lacking a C1B domain (ΔC1B), PKCγ lacking a C1A domain (ΔC1A), or the indicated SCA14 mutants. COS7 cells were treated with the indicated concentrations of PDBu for 24 hours prior to lysis. Endogenous expression of vinculin was also probed as a loading control. N = three independent experiments. *, phosphorylated species; -, unphosphorylated species. (C) Quantification of percent phosphorylation of total PKC as a function of PDBu concentration. Data are mean + S.E.M. (D) Quantification of total levels of PKC with 1000 nM PDBu shown as a percentage of initial levels of PKC (0 nM) and represents mean ± S.E.M. WT levels after 24 hours with 1000 nM PDBu indicated (grey dashed line). *P < 0.05 by Welch’s t-test.