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. 2023 Jan 25;19(1):e1010610. doi: 10.1371/journal.pgen.1010610

Fig 3. Complex V knockdown activates the Integrated Stress Response.

Fig 3

(a, b) Representative images of 2–3 day old CVα KD (a), and CVα and ATF4 KD (b) germaria. (c) Schematic of the Integrated Stress Response (ISR). Phosphorylation of eIF2α activates the ISR by inhibiting global cap-dependent translation and selectively upregulating the translation of the transcription factor, ATF4. ATF4 upregulates the transcription of stress response genes including 4E-BP and Atf3. (d, e) Representative images of 2–3 day old Control (mCherry) (d) and CVα (e) KD germaria. (f) Frequency of GSCs with high phosphorylated eIF2α relative to surrounding somatic cells from (d, e). (g, h) Representative images of 1-day old Control (mCherry) (g) and CVα (h) KD germaria incubated with O-propargyl-puromycin (OPP) to visualize protein synthesis. (i) Intensity of fluorescently derivatized OPP in GSCs relative to follicle cells of Control (mCherry) KD (n = 31) and CVα KD (n = 40). Data are the mean ± s.d. and an unpaired t-test was used for statistical analysis. (j, k) Representative images of less than one week old Control (j) and CVα (k) KD germaria expressing ATF4 reporter, 4E-BPintron-dsRed. (l) Frequency of dsRed-positive GSCs in (g, h). For (f, l) number of GSC analyzed and P-value (Fisher’s exact test) are given above bars. For (a, b) white-dashed lines demark the germline, and for (d-e, g-h, j-k) white-dashed line demark GSCs. All RNAi were driven by nos-GAL4. For exact genotypes see S2 Table.