Fig 5. MrcA disruption or ITPR3 depletion decreases pT853-MYPT1 and pS19-MLC2 recruitment to chlamydial inclusion.

(A) Hela cells were infected with C. trachomatis L2 or mrcA::bla L2. ITPR3 was depleted from Hela cells using siRNA and infected with C. trachomatis L2. At 24 hrs post infection, cells were fixed and stained with anti-pY419-Src (red), anti-pY464-MLCK (red), anti-pT853-MYPT1 (red), anti-pS19-MLC2 (red), anti-MLC2 (red) (all red staining of microdomains is indicated by arrowheads), and anti-MOMP (green). Bar = 10 μm. (B) Immunoblots of L2 and mrcA::bla infected HeLa cells or ITPR3-depleted, L2 infected HeLa cells probed with anti-pT853-MYPT1, anti-pY464-MLCK, anti-MLC2, and anti-pS19-MLC2, with anti-GAPDH as a loading control.