Figure EV4. Inhibition of fat body endocytosis causes muscle detachment, increasing Vkg expression in the fat body improves muscle integrity in tumour‐bearing animals.

• A, A'
  Antibody staining against GFP without permeabilisation can detect Vkg‐GFP.
• B, C
  Nidogen staining in muscles of animals where mCherry ^RNAi or shi ^TS was expressed in the fat body via CG‐GAL4.
• D
  Quantification of muscle nidogen staining in (B, C). mCherry ^RNAi (n = 23), shi ^TS (n = 24).
• E, F
  Muscle fillets stained with phalloidin (Actin) from animals where mCherry ^RNAi or shi ^TS was expressed in the fat body via CG‐GAL4.
• G
  Quantification of normalised muscle detachment in (E, F). mCherry ^RNAi (n = 8), shi ^TS (n = 9).
• H, I
  Muscle fillets stained with phalloidin (Actin) from QRas ^V12 scrib ^RNAi tumour‐bearing animals, where mCherry ^RNAi or UAS‐VkgGFP;UAS‐Cg25C‐GFP was expressed in the fat body (r4‐GAL4). Day 6 muscles used here.
• J
  Quantification of normalised muscle detachment in (H, I). w ¹¹¹⁸ (n = 4), QRas ^V12 scrib ^RNAi ;r4>mCherry ^RNAi (n = 9), QRas ^V12 scrib ^RNAi ;r4>UAS‐VkgGFP;UAS‐Cg25C‐GFP (n = 6).
• K, L
  Fat body staining for Rab10 in w ¹¹¹⁸ and QRas ^V12 scrib ^RNAi tumour‐bearing animals.
• M
  Quantification of normalised Rab10 levels in (K, L). w ¹¹¹⁸ (n = 15), QRas ^V12 scrib ^RNAi (n = 14).
• N, O
  Fat body staining for SPARC in w ¹¹¹⁸ and QRas ^V12 scrib ^RNAi tumour‐bearing animals.
• P
  Quantification of normalised SPARC levels in (N, O). w ¹¹¹⁸ (n = 12), QRas ^V12 scrib ^RNAi (n = 13).

Data information: Scale bar is 500 μm for muscle fillets (E, F, H, I) 50 μm for muscle Nidogen and GFP staining (A, A′, B, C) and 50 μm for fat body staining (K, L, N, O). Muscles in Shi ^TS animals were dissected at day 5 ALH, muscles and fat body in tumour bearing animals were dissected at day 6 after tumour induction. Graphs are represented as Mean ± SEM, n = the number of samples. (*) P < 0.05 (**) P < 0.01, (****) P < 0.0001. For experiments with two genotypes, two‐tailed unpaired student's t‐tests were used to test for significant differences. The Welch's correction was applied in cases of unequal variances. For experiments with more than two genotypes, significant differences between specific genotypes were tested using a one‐way ANOVA and a subsequent Šidák post‐hoc test.