Fig. 3

MO-mediated smc3 knockdown results in reduced regenerate length, segment length and cell proliferation. (A) Representative images of uninjected (UN), smc3 MO-injected (MO1) and Std-MO injected fins. Total regenerate length was calculated by measuring the distance between the amputation plane (indicated by a solid black line) to the distal end of the third fin ray (black arrows indicate the length measured). (B) Graph shows the significant reduction (indicated by *) of regenerate length in smc3 knockdown fins (for both MO1 and MO2) compared to the Std-MO injected fins using the percent similarity method. (C) Representative images of calcein stained fins of uninjected (UN), smc3 MO injected (MO1) and Std-MO injected fins. Segment length was calculated by measuring the distance between first two joints in the 3rd fin ray (black arrows indicates the length measured). Higher magnification images of segments are shown with joints indicated by white arrowheads. (D) Graph shows that significant reduction (indicated by *) of segment length in smc3 knockdown (for both MO1 and MO2) compared to Std-MO injected fins using the percent similarity method. (E) Representative images of H3P-positive cells in uninjected (UN), smc3 MO injected (MO1) and Std-MO injected fins. Measurements were taken from the distal most 250 μm of the 3rd ray. White brackets mark the defined area and n represents the number of H3P-positive cells in that area. Arrows identify H3P-positive cells. (F) Graph shows the significant reduction (indicated by *) in the number of H3P-positive cells in smc3 knockdown (for both MO1 and MO2) compared to Std-MO injected fins using the percent similarity method. For each experiment n=8 fins per trial and three independent trials were performed. *P<0.05, two tailed unpaired Student's t-test. Data are mean±s.e.m. Scale bars: 50 µm in A; 100 µm in E.