Journal: PLoS Biology

Article Title: Distinct Roles for Arp2/3 Regulators in Actin Assembly and Endocytosis

doi: 10.1371/journal.pbio.0060001

Figure Lengend Snippet: (A–G) Null mutants lacking one or both type-I myosin. Cells express Sla2-GFP (A–D) or Abp1-GFP (E–G). (A) MSD plots for wild-type and myo5 Δ patches aligned at the start (left) or end (right) of their lifetimes. The curves on the left are truncated at the median lifetime. (B) Percentage of patches that leave the origin. Mean of values for three segregants is shown. (C) Average time at the origin, from the appearance of a patch until it moved away or disappeared. (D) Phase II movement only. For each patch, data prior to movement were removed. (E) MSD plots for wild-type and mutant patches aligned at the start (left) or end (right) of their lifetimes. On the left, plots are truncated at the median lifetime or 16 s, whichever was less. (F) Percentage of patches that leave the origin, as in (B). (G) Average time at the origin, as in (C). (H–M) Mutants lacking Arp2/3-binding regions of one or both type-I myosins, termed myo3 Δ acidic and myo5 Δ acidic . The panels are similar to those above, with cells expressing Sla2-GFP (H–J) or Abp1-GFP (K–M). (H and K) MSD plots for wild-type or mutant patches aligned at the start (left) or end (right) of their lifetimes. The curves on the left are truncated at the median lifetime. (I and L) Percentage of patches that leave the origin. Mean of values for three segregants is shown. (J and M) Average time at the origin, from the appearance of a patch until it moved away or disappeared. (N–S) Mutants lacking Arp2/3 binding regions of WASp/Las17 and type-I myosins. Cells express Sla2-GFP (N–P) or Abp1-GFP (Q–S). (N and Q) MSD plots for wild-type or mutant patches aligned at the start (left) or end (right) of their lifetimes. The curves on the left are truncated at the median lifetime. (O and P) Percentage of patches that leave the origin. Mean of values for three segregants is shown. (R and S) Average time at the origin, from the appearance of a patch until it moved away or disappeared. Strain numbers and number of patches: (A–C) Wild-type, YJC4787–9; 94, 79, and 90. myo5 Δ, YJC4784–6; 103, 79, and 69. (D) Strains as in (A–C). Numbers of patches: wild-type – 88, 69, 85. myo5 Δ – 75, 40, 36. (E–G) Wild-type, YJC4815–7; 121, 110, and 130. myo3 Δ, YJC4409–11; 111, 115, and 105. myo5 Δ, YJC4813–4; 120 and 99. myo3 Δ myo5 Δ, YJC4770–2; 143, 141, and 126. (H–J) Wild-type, YJC5566–8; 71, 107, and 83. myo3 Δ acidic , YJC5563–5; 71, 75, and 101. myo5 Δ acidic , YJC5554–6; 86, 71, and 62. myo3 Δ acidic myo5 Δ acidic , YJC5557–9; 83, 65, and 83. (K–M) Wild-type, YJC4848 and 50; 116 and 87. myo3 Δ acidic , YJC4843–5; 119, 99, and 106. myo5 Δ acidic , YJC4840–2; 102, 111, and 99. myo3 Δ acidic myo5 Δ acidic , YJC4837–9; 96, 93, and 100. (N–P) Wild-type, YJC5566–8; 71, 107, and 83. las17 Δ acidic , YJC5551–3; 84, 64, and 69. las17 Δ acidic myo3 Δ acidic , YJC5560–2; 89, 71, and 68. las17 Δ acidic myo5 Δ acidic , YJC5548–50; 51, 66, and 57. las17 Δ acidic myo3 Δ acidic myo5 Δ acidic , YJC5545–7; 43, 53, and 54. (Q–S) las17 Δ acidic , YJC5208–10; 95, 76, and 89. las17 Δ acidic myo3 Δ acidic , YJC5438–40; 111, 104, and 109. las17 Δ acidic myo5 Δ acidic , YJC5441–3; 98, 80, and 84. las17 Δ acidic myo3 Δ acidic myo5 Δ acidic , YJC5205–7; 71, 79, and 99.

Article Snippet: Two color images were collected sequentially using a LMM5 Laser Merge Module (Spectral Applied Research), a multipass dichroic mirror, and emission filters in a Lambda 10–3 high-speed filter wheel (Sutter Instruments).

Techniques: Mutagenesis, Binding Assay, Expressing

Journal: PLoS Biology

Article Title: Distinct Roles for Arp2/3 Regulators in Actin Assembly and Endocytosis

doi: 10.1371/journal.pbio.0060001

Figure Lengend Snippet: Cells express Sla2-GFP (A–C) or Abp1-GFP (D–F). ( A and D) MSD plots for wild-type and mutant patches aligned at the start (left) or end (right) of their lifetimes. The curves on the left are truncated at the median lifetime. (B and E) Percentage of patches that leave the origin. Mean of values for three segregants is shown. (C and F) Average time at the origin, from the appearance of a patch until it moved away or disappeared. Strain numbers and numbers of patches: (A–C) Wild-type, YJC5707 - 9; 66, 84, and 69. las17 Δ acidic , YJC5713–5; 100, 95, and 104. pan1 Δ acidic , YJC5710–2; 103, 76, and 74. las17 Δ acidic pan1 Δ acidic , YJC5716–8; 100, 86, and 91. (D–F) Wild-type, YJC5719–22; 99, 100, 114, and 98. las17 Δ acidic , YJC5726, 8 & 9; 95, 88, and 84. pan1 Δ acidic , YJC5723–5; 102, 94, and 139. las17 Δ acidic pan1 Δ acidic , YJC5730–3; 77, 70, 50, and 50. Error bars are ± standard error.

Article Snippet: Two color images were collected sequentially using a LMM5 Laser Merge Module (Spectral Applied Research), a multipass dichroic mirror, and emission filters in a Lambda 10–3 high-speed filter wheel (Sutter Instruments).

Techniques: Mutagenesis