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. 2011 Jul 11;121(8):3189–3205. doi: 10.1172/JCI42975

Figure 2. Effects of LCRMP-1 on filopodia formation.

Figure 2

(A and B) Localization of (A) endogenous LCRMP-1 (green), (B) exogenous GFP-LCRMP-1 (green), and actin (red) by immunofluorescent staining. (A) Preimmune serum and (B) CL1-0/CL1-5 mock were used as controls. LCRMP-1 shares some common compartments with actin in its distribution, especially in lamellipodia and filopodia regions. Arrows indicate presence of LCRMP-1 in the filopodia region. (B) In addition, exogenous GFP–LCRMP-1 promotes filopodia formation in both CL1-0 and CL1-5 cells. Numbers of filopodia were counted (n = 20 cells per group; original magnification, ×1,000 [A, top 2 and the bottom rows, and B, top and bottom row]; ×4,000 [A, third row, and B, middle row]). (C) Schematic of GFP-tagged LCRMP-1 N-terminal deletion mutants (LCRMP-1, LCRMP-1–P28AQ30A, LCRMP-1–R29AK31A, LCRMP-1Δ22, LCRMP-1Δ72, LCRMP-1Δ105, LCRMP-1Δ127, and CRMP-1; the double asterisks indicate the 2 mutation sites of each point mutation). (D) CL1-0 cells were transfected with indicated GFP-tagged LCRMP-1 N-terminal deletion constructs and actin stained with rhodamine-conjugated phalloidin (red). Number of filopodia were counted (n = 20 cells per group; original magnification, ×1,000). Data are presented as mean ± SEM, and P values were calculated by 2-sided Student’s t test.