Requirements of CID, CAL1, and CENP-C for their recruitment and stability in meiosis I. (A) Pupal testes stained with anti-CENP-C (green) and anti-CID (magenta) antibodies, and DAPI (blue). Representative images of early prophase I (S1 stage) cells from each of the genetic lines (bam-GAL4, CID RNAi, CAL1 RNAi, and CENP-C RNAi). Scale bars = 3 µm. (B) Quantitation showing the relative fluorescent intensity per nucleus of CID at S1 in early prophase I. n = 46, 37, 39, 34. Here, and throughout this figure, n represents the number of cells analyzed from at least three independent replicates; data were checked for normality using the Shapiro–Wilk tests and analyzed using either Welch’s or Mann–Whitney tests. Error bars = SEM. (C) Quantitation showing the relative fluorescent intensity per nucleus of CENP-C at S1 in early prophase I. n = 57, 47, 30, 31. Error bars = SEM. (D) Early prophase I (S1 stage) cells stained with anti-CAL1 (green) and anti-CID (magenta) antibodies, and DAPI (blue). Scale bars = 3 µm. (E) Quantitation showing the relative fluorescent intensity per nucleus of the histone chaperone CAL1 at S1 in early prophase I. n = 46, 37, 55, 61. Error bars = SEM. (F) Prometaphase I cells stained with anti-CENP-C (green) and anti-CID (magenta) antibodies, and DAPI (blue). Each panel shows one nucleus, visible as three to four chromosome territories at prometaphase I. Scale bars = 5 µm. (G) Quantitation showing the relative fluorescent intensity per nucleus of CID at prometaphase I. n = 42, 33, 45, 27. Error bars = SEM. (H) Quantitation showing the relative fluorescent intensity per nucleus of CENP-C at prometaphase I. n = 50, 50, 46, 49. Error bars = SEM. (I) Quantitation showing the fluorescent intensity of CID per nucleus from 8CC, early prophase I (S1), late prophase I (S6), and the spermatid stages T1 and T5 for bam-GAL4 (shown in gray), CID RNAi (red), CAL1 RNAi (green), and CENP-C RNAi (blue). Fold changes in the first phase of CID assembly are annotated between S1 and S6 for each line. Similarly, fold changes are shown between T1 and T5 for the second phase of assembly. Significant percent reductions in CID intensities relative to the same cell stage from the bam-GAL4 control are indicated for each RNAi line. n = 30–50. Error bars = SEM. (J) Model depicting the interdependencies for CID, CAL1, and CENP-C recruitment during meiosis I. Arrows represent a complete dependency between these proteins for their centromeric recruitment, with the color indicating the direction of this dependency. Black lines indicate incomplete dependency.