Figure 6. Hindsight regulates Mmp2 expression in stage-14 follicle cells.

(A–H) Response of egg chambers isolated according to FC1 (A–D) or FC2 (E–H) to ionomycin-induced rupture in three hours. (A–C and E–G) Representative images show control (A and E) and hnt^RNAi (B–C and F–G) egg chambers after the culture. Bright field images of the egg chambers are shown in blue, and white arrowheads mark ruptured egg chambers. Quantification of rupture response to OA or to ionomycin is shown in (D and H). (I–K) Representative images show Mmp2::GFP expression (green in I-K and white in I’-K’) in control (I–I’) or hnt^RNAi (J–K’) egg chambers with FC2 Gal4 (Red). Nuclei are labeled with DAPI and shown in blue (I–K). Arrowheads point to posterior follicle cells, and oocytes are outlined in cyan (I’–K’). (L–M) Quantification of Mmp2::GFP expression in control and hnt^RNAi egg chambers using FC1 (L) or FC2 (M) Gal4. (N–O) Quantification of Mmp2 mRNA levels in hnt^RNAi egg chambers with FC1 (N) or FC2 (O) Gal4. *p<0.05, **p<0.01, ***p<0.001.

Figure 6—figure supplement 1. Measurement of intracellular Ca2²⁺in follicle cells after ionomycin or octopamine stimulation.

(A–D) Intracellular Ca²⁺ depicted by GCaMP5G (green in A-C’) increase in response to ionomycin stimulation in both control (A–A’) and hnt^RNAi (B–C’) egg chambers with FC2 Gal4, although hnt^RNAi egg chambers show slightly weaker response. The frames with peak GCaMP5G signal after ionomycin stimulation are shown in A’-C’. Quantification of intracellular Ca²⁺ level (ΔF/F0) is shown in D and the number of egg chambers analyzed is noted at the end of each trace. (E–H) Intracellular Ca²⁺ depicted by GCaMP5G (green in E-G’) increase in control egg chambers (E–E’) after OA stimulation but does not in hnt^RNAi egg chambers with FC2 Gal4 (F–G’). The frames with peak GCaMP5G signal after OA stimulation are shown in E’-G’. Quantification of intracellular Ca²⁺ level (ΔF/F0) is shown in H and the number of egg chambers analyzed is noted at the end of each trace. Also see Videos 1–6.

Figure 6—figure supplement 2. hnt depletion disrupts Mmp2 expression in posterior follicle cells.

Mmp2::GFP (green in A-C and white in A’-C’) is highly expressed in posterior follicle cells of control stage-14 egg chambers (A–A’), but is weakly detected in posterior follicle cells of hnt^RNAi egg chambers (B–C’) with FC1 Gal4. The oocyte is outlined in cyan (A’–C’), and nuclei are labeled with DAPI and shown in blue (A–C).