Figure 7.

Cx43 specific inhibitor α-CT1 reduces intercellular communication in PKCμ inhibited/depleted HaCaT cells

(A) Amino acid sequence of peptides α-CT1, reverse inactive peptide (α-CT1 RIS), and antennapedia control (ANTP).

(B) HaCaT cells were scratched with a pipette tip followed by treatment with PMA (10 nM) with or without CRT (5 μM), in the absence or presence of ANTP, α-CT1 RIS, or α-CT1 peptides (each at 100 μM) for 16 h. Cell lysates were probed by western blotting for pCx43-S368 Cx43, pPKCμ-S916, PKCμ, and tubulin.

(C) Relative pCx43-S368 levels in treated samples compared to DMSO control based on densitometric analysis from three replicate experiments as shown in (A).

(D) Representative fluorescent microscopic images of SL/DT assay in HaCaT cells treatment with or without CRT (5 μM), in the absence or presence of ANTP, α-CT1 RIS, or α-CT1 peptides (each at 100 μM) for 6 h followed by PMA (10 nM) for 30 min. Scale bar = 100 μM.

(E) Quantification of dye migration area from three replicate experiments as shown in (D).

(F) Representative fluorescent microscopic images of SL/DT assay in HaCaT cells transduced with control shRNA (shGFP) or an shRNA directed against PKCμ and in the absence or presence of ANTP, α-CT1 RIS, or α-CT1 peptides (each at 100 μM) for 6 h followed by PMA (10nM) for 30 min. Scale bar = 100 μm.

(G and H) Quantification of dye migration area from three replicate experiments as shown in (F) for shGFP (G) or shPKCμ.

(H) All calculations are based on three replicates −/+ S.D. ∗∗p < 0.01, ∗∗∗p < 0.001 (Student’s t test).