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. 2023 Jun 14;6(9):e202301920. doi: 10.26508/lsa.202301920

Figure 5. PIP4K regulates a non-endosomal PI3P pool in Drosophila salivary glands.

Figure 5.

(A) Graph representing average cell size measurement (in μm3) as mean ± S.E.M. of salivary glands from wandering third instar larvae of AB1/+; dPIP4K29 (n = 9), AB1>PI3K59F RNAi; dPIP4K29 (n = 9). Sample size is represented on individual bars. Unpaired t test with Welch correction showed P-value < 0.0001. (B) Graph representing normalised PI3P levels which is the peak area of GroPI3P/peak area of GroPI4P normalised to organic phosphate value of total lipid extracts as mean ± S.E.M. of Act5C/+; dPIP4K29 (green) or Act5C> PI3K59F RNAi; dPIP4K29 (magenta). Biological samples = 5, where each sample was made from five third instar wandering larvae. Unpaired t test with Welch correction showed P-value = 0.011. (C) (i) Representative confocal z-projections depicting a subpopulation of early endosomal compartment using 2XFYVE-mCherry in the salivary glands from wandering third instar larvae of AB1> 2XFYVE::mCherry (Control) and AB1> 2XFYVE::mCherry; dPIP4K29. Scale bar indicated at 20 μm. (ii) Graph representing 2XFYVE punctae measurement in the salivary glands from wandering third instar larvae of AB1> 2XFYVE::mCherry (N = 8, n = 40) and AB1>2XFYVE::mCherry; dPIP4K29 (N = 8, n = 40). Unpaired t test with Welch correction showed P-value = 0.4057. (D) Graph representing average cell size measurement (in μm3) as mean ± S.E.M. of salivary glands from wandering third instar larvae of AB1/+; dPIP4K29 (n = 8), AB1>dPIP4K2XFYVE; dPIP4K29 (n = 8). Sample size is represented on individual bars. Unpaired t test with Welch correction showed P-value = 0.171. (E) qPCR measurements for mRNA levels of UVRAG from either da/+ (green) or da> UVRAG RNAi (magenta). Unpaired t test showed P-value = 0.0046. (n = 6, where each n is derived from five third instar wandering larvae). (F) Graph representing average cell size measurement as mean ± S.E.M. of salivary glands from wandering third instar larvae of AB1/+; dPIP4K29 (n = 12) and AB1> UVRAG RNAi; dPIP4K29 (n = 12). unpaired t test with Welch correction showed P-value = 0.4306. (G) (i) Representative confocal z-projections depicting 2XFYVE punctae using 2XFYVE::mCherry in the salivary glands from the genotypes a. AB1> 2XFYVE::mCherry (Control) b. AB1> 2XFYVE::mCherry; UVRAG RNAi. (ii) Graph representing measurement of 2XFYVE punctae numbers per unit cell area of the salivary glands from wandering third instar larvae of AB1> 2XFYVE::mCherry (N = 3, n = 11), AB1> 2XFYVE::mCherry; UVRAG RNAi (N = 3,n = 11).