Fig. 5
Knocking down His3 or His4 recapitulates the mxc phenotypes. A, B Quantitative RT-PCR results revealed that transcription levels of canonical histone genes were decreased in mxc RNAi (A) and mxc16a − 1 (B) larval brains. The data are plotted as mean ± SD. ****p < 0.0001, ***p < 0.001 and **p < 0.01 using a Student’s t test, p = 0.0003, 0.0002, 7.758E–05, 0.0001, 0.0014 in (A) and 0.0006, 0.0003, 0.0007, 0.0013, 4.019E-05 in (B), respectively. Both of the numbers of repetitions N = 3. C The western blot results shown that the protein levels of His3 are reduced in mxc knockdown larval brain compared to wt. D–F’ Third-instar larval brains of wt (D–D’), His3 RNAi (E–E’), and His4 RNAi (F–F’), labeled by anti-Pros (red) and anti-Dpn (green). Pros (arrowheads) was detected in the nucleus of NBs in both His3 (E) and His4 (F) RNAi lines. Scale bars: 20 μm. G Statistical data of nuclear Pros in the NBs in wt, His3 RNAi, His4 RNAi, and mxc RNAi larval central brain. The data are plotted as mean ± SD. **p < 0.01 and ***p < 0.001 using a Student’s t test, p = 0.0037 and 9.958E-04, respectively. Numbers of NBs counted N = 360, N = 740, N = 151, N = 85, N = 740, respectively. H–J’ Type-I NB FLP-out clones of wt (H–H’), His3 RNAi (I–I’) and His4 RNAi J–J’, labeled by anti-Dpn (red) and anti-GFP (green). In wt clones (H), Dpn was only expressed in the NBs. In both His3 and His4 RNAi clones (I, J), Dpn was detected in the GMCs (arrowheads). Scale bars: 10 μm. K Statistical analysis of the numbers of Dpn+ cells in the larval central brains of wt, His3 RNAi, His4 RNAi and mxc RNAi lines. The data are plotted as mean ± SD. **p < 0.01 using a Student’s t test, p = 0.0075 and 0.0070, respectively. Numbers of brain lobes counted N = 6, N = 5, n = 4, N = 4, respectively