Journal: Cell Death & Disease

Article Title: Role of autophagy in modulating post-maturation aging of mouse oocytes

doi: 10.1038/s41419-018-0368-5

Figure Lengend Snippet: Graphs a and d show percentages of activated oocytes. While a shows freshly collected control (Ctrl) oocytes and oocytes aging for 12 h in CZB + MG132 (C + M), CZB or FCM, d shows oocytes aging for different times in FCM. Each treatment was repeated 3–4 times with each replicate containing about 30 oocytes. Graphs b and c show LC3-II quantification by western blotting in oocytes aging for 12 h in different media. While b shows LC3-II contents (LC3-II/tubulin ratio), c shows the LC3-II/LC3-I ratio. Each treatment was repeated three times with each replicate containing about 200–250 oocytes. Graph e shows relative levels of active caspase-3 in oocytes aging in FCM for different times as detected by western blotting. Each treatment was repeated three times with each replicate containing about 300-400 oocytes. Graphs f , g and h show LC3-II contents (LC3-II/GAPDH ratio), LC3-II/LC3-I ratio and p62 (p62/GAPDH ratio), respectively, in oocytes aging in FCM for different times. Each treatment was repeated three times with each replicate containing about 200–250 oocytes. a – d Values with a different letter above bars differ significantly ( P < 0.05). Micrographs i , j and k are confocal images showing autophagosome (LC3-II puncta) distribution in oocytes aging in FCM for 0, 12 and 18 h, respectively. These are the merged pictures with chromosomes and LC3-II puncta pseudo colored blue and red, respectively. The bar is 15 µm and applies to all images

Article Snippet: The primary antibodies used included rabbit anti-LC3 antibody (4108, Cell Signaling, USA), rabbit anti-active caspase-3 polyclonal antibodies (ab13847, Abcam Co. Ltd, Cambridge, MA, USA), rabbit anti-P62 antibody (5114 S, Cell Signaling), mouse anti-β-tubulin monoclonal antibodies (05-661, Merck Millipore, China), and mouse anti-GAPDH monoclonal antibody (CW0100A, CWBIO, China).

Techniques: Control, Western Blot

Journal: Cell Death & Disease

Article Title: Role of autophagy in modulating post-maturation aging of mouse oocytes

doi: 10.1038/s41419-018-0368-5

Figure Lengend Snippet: Oocytes that had aged for 12 h in FCM or FCM containing 10 nM Rapamycin, 10 mM LiCl or 5 mM 3-MA were cultured for 24 h in CZB medium before examination for morphology of spindles and chromosomes. In the confocal images, DNA and α-tubulin were pseudo-colored blue and green, respectively. Bar is 10 µm and applies to all images. Image a shows an oocyte with a tine-pole spindle and chromosomes congressed on the metaphase plate (T/C), image b shows an oocyte with a barrel-shaped spindle and congressed chromosomes (B/C), image c shows an oocyte with a disintegrated spindle and congressed chromosomes (D/C), and image d shows an oocyte with a disintegrated spindle and scattered chromosomes (D/S). Graph e shows percentages of oocytes with different spindle/chromosome configurations following oocytes were aged in different media. Each treatment was repeated 3 times with each replicate containing about 20 oocytes. a – c Values with different letters above bars differ significantly ( P < 0.05) within spindle/chromosome morphologies

Article Snippet: The primary antibodies used included rabbit anti-LC3 antibody (4108, Cell Signaling, USA), rabbit anti-active caspase-3 polyclonal antibodies (ab13847, Abcam Co. Ltd, Cambridge, MA, USA), rabbit anti-P62 antibody (5114 S, Cell Signaling), mouse anti-β-tubulin monoclonal antibodies (05-661, Merck Millipore, China), and mouse anti-GAPDH monoclonal antibody (CW0100A, CWBIO, China).

Techniques: Cell Culture

Journal: Research Square

Article Title: Herpesviruses mimic zygotic genome activation to promote viral replication

doi: 10.21203/rs.3.rs-3125635/v1

Figure Lengend Snippet: Antibodies

Article Snippet: B-Tubulin antibody , mouse monoclonal , Genescript , A01717-40.

Techniques: Purification

Journal: Scientific Reports

Article Title: Long noncoding RNA lnc-RI is a new regulator of mitosis via targeting miRNA-210-3p to release PLK1 mRNA activity

doi: 10.1038/srep25385

Figure Lengend Snippet: ( a,b ) Knockdown of lnc-RI delayed M-phase progression. HeLa cells were transfected with lnc-RI RNAi-2 or Ctrl RNAi. At 12 h after transfection, the cells were treated with nocodazole (100 ng/mL) for 16 h. Mitotic cells were harvested using the mitotic shake-off method. Cell cycle profile was performed at the indicated time points after the cells were released. ( c ) The knockdown of lnc-RI induced aberrant mitotic spindle in HeLa cells. HeLa cells were transfected with lnc-RI RNAi-1, lnc-RI RNAi-2, and Ctrl RNAi and stained with anti-β-tubulin antibody and DAPI at 48 h after transfection. ( d ) The percentages of HeLa cells harbouring aberrant spindles were calculated in 200 randomly selected mitotic cells 48 h after siRNA transfection.

Article Snippet: The following antibodies were used: rabbit anti-phospho-histone H3 (S10) polyclonal antibody (A301-844A, Bethyl, USA); mouse anti-β-tubulin (D-10) monoclonal antibody (ZS-5274, ZSGB-BIO, China); mouse anti-PLK1 (F-8) monoclonal antibody (sc-17783, Santa Cruz, USA); mouse anti-β-actin monoclonal antibody (60008-1-lg, Proteintech, USA); HRP-conjugated anti-mouse IgG (074-1806, KPL, USA); FITC-conjugated anti-rabbit IgG (02-15-06, KPL,USA); and FITC-conjugated anti-mouse IgG (02-18-06, KPL,USA).

Techniques: Knockdown, Transfection, Staining