FMNL2 regulates actin for endoplasmic reticulum and mitochondria distribution in oocyte meiosis
- College of Animal Science and Technology, Nanjing Agricultural University, China
- College of Veterinary Medicine, Northwest A&F University, China
- Fertility Preservation Lab, Reproductive Medicine Center, Guangdong Second Provincial General Hospital, China
Figures
Figure 1 with 1 supplement
Expression and subcellular localization of FMNL2 during mouse oocyte meiosis.
(A) Western blotting results of FMNL2 protein expression at different stages. FMNL2 expressed at the germinal vesicle (GV), metaphase I (MI), and metaphase II (MII) stages. (B) Subcellular …
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Figure 1—source data 1
The original files of the full raw unedited blots in Figure 1.
- https://cdn.elifesciences.org/articles/92732/elife-92732-fig1-data1-v1.zip
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Figure 1—source data 2
The figure with the uncropped blots with the labeled bands.
- https://cdn.elifesciences.org/articles/92732/elife-92732-fig1-data2-v1.zip
Figure 1—figure supplement 1
Localization of FMNL2 in the different stages of porcine oocyte maturation.
FMNL2 colocalized with actin in porcine oocytes. Green, FMNL2; red, actin; blue, DNA. Bar = 20 μm.
Figure 2
Knockdown of FMNL2 affects first polar body extrusion and asymmetric division.
(A) Western blot analysis for FMNL2 expression in the FMNL2-KD group and control group. Relative intensity of FMNL2 and tubulin was assessed by densitometry. (B) Brightfield images of control …
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Figure 2—source data 1
The original files of the full raw unedited blots in Figure 2.
- https://cdn.elifesciences.org/articles/92732/elife-92732-fig2-data1-v1.zip
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Figure 2—source data 2
The figure with the uncropped blots with the labeled bands.
- https://cdn.elifesciences.org/articles/92732/elife-92732-fig2-data2-v1.zip
Figure 3 with 1 supplement
Knockdown of FMNL2 disrupts spindle localization during mouse oocyte meiosis.
(A) Time-lapse microscopy showed that spindle migration failed after FMNL2-KD. Green, tubulin-EGFP. Bar = 10 μm. (B) Representative images and the proportion of spindle migration after 9.5 hr of …
Figure 3—figure supplement 1
The spindle positioning after FMNL2 antibody injection in porcine oocytes.
We defined oocyte diameter as D, and the length of spindle to the cortex as L. The ratio of L/D increased significantly in FMNL2 antibody injection group. Green, tubulin; blue, DNA. Bar = 20 μm. The …
Figure 4 with 1 supplement
Knockdown of FMNL2 disrupts actin assembly during mouse oocyte meiosis.
(A, B) Representative images of actin distribution at the oocyte cortex and the fluorescent intensities in the control group and FMNL2-KD group (p > 0.1). White, actin; green, tubulin; blue, DNA. …
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Figure 4—source data 1
The original files of the full raw unedited blots in Figure 4.
- https://cdn.elifesciences.org/articles/92732/elife-92732-fig4-data1-v1.zip
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Figure 4—source data 2
The figure with the uncropped blots with the labeled bands.
- https://cdn.elifesciences.org/articles/92732/elife-92732-fig4-data2-v1.zip
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Figure 4—source data 3
The original file of mass spectrometry for the protein summary.
- https://cdn.elifesciences.org/articles/92732/elife-92732-fig4-data3-v1.zip
Figure 4—figure supplement 1
The actin intensity in the cytoplasm of porcine oocytes.
The intensity of cytoplasmic actin decreased after FMNL2 antibody injection.Red, actin; blue, DNA. Bar = 20 μm. Control (n = 42), FMNL2‐antibody (n = 40). The error bars are representing the …
Figure 5 with 1 supplement
FMNL2 regulates endoplasmic reticulum (ER) distribution during mouse oocytes maturation.
(A) Mass spectrometry results showed that FMNL2 was associated with ER-related proteins. (B) Co-IP results showed that FMNL2 was correlated with INF2. (C) Representative images of ER distribution in …
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Figure 5—source data 1
The original files of the full raw unedited blots in Figure 5.
- https://cdn.elifesciences.org/articles/92732/elife-92732-fig5-data1-v1.zip
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Figure 5—source data 2
The figure with the uncropped blots with the labeled bands.
- https://cdn.elifesciences.org/articles/92732/elife-92732-fig5-data2-v1.zip
Figure 5—figure supplement 1
The endoplasmic reticulum (ER) distribution in porcine oocytes.
The rate of abnormal ER increased after FMNL2 antibody injection. Blue, ER. Bar = 20 μm. The error bars are representing the mean ± SEM. The P‐values were calculated using Student's t‐test. *p < 0.05.
Figure 6 with 1 supplement
FMNL2 regulates mitochondrial distribution during mouse oocytes maturation.
(A) Mass spectrometry results showed that FMNL2 was related to many mitochondria-related proteins. (B) Representative images of mitochondrial distribution in the oocyte cytoplasm in the control …
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Figure 6—source data 1
The original files of the full raw unedited blots in Figure 6.
- https://cdn.elifesciences.org/articles/92732/elife-92732-fig6-data1-v1.zip
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Figure 6—source data 2
The figure with the uncropped blots with the labeled bands.
- https://cdn.elifesciences.org/articles/92732/elife-92732-fig6-data2-v1.zip
Figure 6—figure supplement 1
The mitochondria distribution in porcine oocytes.
The rate of abnormal mitochondria increased after FMNL2 antibody injection.Green, mitochondria. Bar = 20 μm. The error bars are representing the mean ± SEM. The P‐values were calculated using …
Figure 7
Diagram of the roles of FMNL2 during oocyte maturation.
FMNL2 associates with Formin2 and Arp2/3 complex for actin assembly, which further regulates spindle migration and INF2/Cofilin-related organelle dynamics during mouse and porcine oocyte maturation.
