Figure S5.
Related toFig. 6. Effect of Lat-A and actin nucleators on PVD branching and F-actin distribution. (A) Schematic showing the experimental design of Lat-A treatment. To obtain synchronized L1 animals and for auxin treatment the protocol in Fig. S2 A was followed. Once the animals reached late L3 stage (16 h after auxin), they were exposed to 100 µM of Lat-A or DMSO in a humidity chamber for 2 h. The animals were then transferred to plates with food and auxin and grown until they reached late L4 stage. (B) Quantification of number of 2° branches for the indicated conditions. n represents number of animals. Error bars represent mean ± 95% CI. P values from one-way ANOVA test followed by Tukey’s test. (C) Images showing actin (Lifeact::mKate2) in the cell body (CB) and proximal anterior primary dendrite in indicated conditions. Scale bar, 5 µm. (D) Box and whisker plot showing quantification of Lifeact::mKate2 fluorescence intensity along a three-pixel-wide line along the cell body (CB) periphery in control and KNL-1 DEG when treated with DMSO or 100 µM Lat-A. n represents the number of animals. Whiskers represent minimum and maximum whilst bar represents first, second, and third quartiles. P values from one-way ANOVA test followed by Tukey’s test. (E and F) Quantification of the number of 2° branches and protrusions for the indicated conditions. n represents the number of animals. Error bars represent mean ± 95% CI. P values from Kruskal–Wallis test followed by Dunn’s test. (G) Images highlighting actin distribution in the cell body (CB) and proximal anterior primary dendrite for the indicated conditions. Scale bar, 5 µm. (H) Box and whisker plot showing quantification of Lifeact::mKate2 fluorescence intensity along a three-pixel wide line along the circumference of the cell body (CB) in control, KNL-1 DEG, and KNL-1 CYK-1 DEG. n represents the number of animals. Whiskers represent minimum and maximum whilst bar represents first, second, and third quartiles. P values from Kruskal–Wallis test followed by Dunn’s test. (I) Box and whisker plot showing quantification of Lifeact::mKate2 fluorescence intensity of a cross section of the 1° dendrite 10 µm anterior of the cell body along a three-pixel-wide line in indicated conditions. Whiskers represent minimum and maximum while bar represents first, second, and third quartiles. P values from one-way ANOVA test followed by Tukey’s test. (J) Schematic and images of the anterior dorsal menorah proximal to the cell body, showing the actin distribution at the 4° dendrites for the indicated conditions. Actin and membrane are labeled using Lifeact::mKate2 and mNeonGreen(mNG)::PH, respectively. Scale bar, 5 µm. (K) Box and whisker plot showing quantification of the Lifeact::mKate2 fluorescence intensity along a two-pixel-wide line positioned along 4° dendrites of the first anterior menorah. n represents the number of animals. Whiskers represent minimum and maximum whilst bar represents first, second, and third quartiles. P values from Kruskal–Wallis test followed by Dunn’s test.

Related to Fig. 6 . Effect of Lat - A and actin nucleators on PVD branching and F-actin distribution. (A) Schematic showing the experimental design of Lat-A treatment. To obtain synchronized L1 animals and for auxin treatment the protocol in Fig. S2 A was followed. Once the animals reached late L3 stage (16 h after auxin), they were exposed to 100 µM of Lat-A or DMSO in a humidity chamber for 2 h. The animals were then transferred to plates with food and auxin and grown until they reached late L4 stage. (B) Quantification of number of 2° branches for the indicated conditions. n represents number of animals. Error bars represent mean ± 95% CI. P values from one-way ANOVA test followed by Tukey’s test. (C) Images showing actin (Lifeact::mKate2) in the cell body (CB) and proximal anterior primary dendrite in indicated conditions. Scale bar, 5 µm. (D) Box and whisker plot showing quantification of Lifeact::mKate2 fluorescence intensity along a three-pixel-wide line along the cell body (CB) periphery in control and KNL-1 DEG when treated with DMSO or 100 µM Lat-A. n represents the number of animals. Whiskers represent minimum and maximum whilst bar represents first, second, and third quartiles. P values from one-way ANOVA test followed by Tukey’s test. (E and F) Quantification of the number of 2° branches and protrusions for the indicated conditions. n represents the number of animals. Error bars represent mean ± 95% CI. P values from Kruskal–Wallis test followed by Dunn’s test. (G) Images highlighting actin distribution in the cell body (CB) and proximal anterior primary dendrite for the indicated conditions. Scale bar, 5 µm. (H) Box and whisker plot showing quantification of Lifeact::mKate2 fluorescence intensity along a three-pixel wide line along the circumference of the cell body (CB) in control, KNL-1 DEG, and KNL-1 CYK-1 DEG. n represents the number of animals. Whiskers represent minimum and maximum whilst bar represents first, second, and third quartiles. P values from Kruskal–Wallis test followed by Dunn’s test. (I) Box and whisker plot showing quantification of Lifeact::mKate2 fluorescence intensity of a cross section of the 1° dendrite 10 µm anterior of the cell body along a three-pixel-wide line in indicated conditions. Whiskers represent minimum and maximum while bar represents first, second, and third quartiles. P values from one-way ANOVA test followed by Tukey’s test. (J) Schematic and images of the anterior dorsal menorah proximal to the cell body, showing the actin distribution at the 4° dendrites for the indicated conditions. Actin and membrane are labeled using Lifeact::mKate2 and mNeonGreen(mNG)::PH, respectively. Scale bar, 5 µm. (K) Box and whisker plot showing quantification of the Lifeact::mKate2 fluorescence intensity along a two-pixel-wide line positioned along 4° dendrites of the first anterior menorah. n represents the number of animals. Whiskers represent minimum and maximum whilst bar represents first, second, and third quartiles. P values from Kruskal–Wallis test followed by Dunn’s test.

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