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: Glia

Article Title: GFAP expression and social deficits in transgenic mice overexpressing human sAPPα

doi: 10.1002/glia.22544

Figure Lengend Snippet: Suppressed β-tubulin III and elevated GFAP in TgsAPPα-derived murine NSCs. A: Representative photomicrographs of WT littermate and TgsAPPα mouse-derived NSCs under differentiating culture conditions (Day 3) showing increased GFAP (green) and decreased β-tubulin III (red) expression by immunofluorescence (IF). B: WB analysis of GFAP and β-tubulin III expression in both NSCs. C: Densitometric analysis revealed increased ratio of GFAP to β-tubulin III in TgsAPPα mouse-derived NSCs compared to WT littermate control. Data presented as mean ± sd. D: Relative concentrations of sAPPα secreted by WT littermate and TgsAPPα mouse-derived NSCs measured by ELISA and expressed as ng of sAPPα/mL media. TgsAPPα mouse NSCs secrete significantly greater concentrations of sAPPα than WT. These results are representative of three independent experiments with each condition triplicated. *** P < 0.001.

Article Snippet: After blocking with 5% milk in 1X TBS, blots were incubated overnight at 4°C with either of the following primary antibodies: mouse monoclonal anti-Aβ 1-17 (6E10, 1:1,000, Covance Research Products); mouse glial fibrillary acidic protein (GFAP) (1:1,000, Cell Signaling Technology); mouse monoclonal anti-β-tubulin (1:1,000, Stem Cell Technologies, Tukwila, WA); rabbit polyclonal anti-APP C terminus (pAb 396, 1:1,000, kindly provided by S. Gandy and H. Steiner); rabbit polyclonal anti-Notch1 (1:1,000, Epitomics, Burlingame, CA); rabbit polyclonal anti-Notch 1 intercelluar domain (NICD, activated Notch 1) (1:500, Abcam, Cambridge, MA); rabbit polyclonal anti-gp130 (1:1,000, Merck Millipore, Darmstadt, Germany); mouse monoclonal anti-β-actin (1:4,000, Sigma-Aldrich).

Techniques: Derivative Assay, Expressing, Immunofluorescence, Enzyme-linked Immunosorbent Assay

Journal: Glia

Article Title: GFAP expression and social deficits in transgenic mice overexpressing human sAPPα

doi: 10.1002/glia.22544

Figure Lengend Snippet: Suppressed β-tubulin III and elevated GFAP in WT murine NSCs treated with recombinant human sAPPα (rhsAPPα). A: Representative photomicrographs of murine NSCs treated with 2 nM rhsAPPα protein under differentiating culture conditions (Day 5) revealed enhanced GFAP (green) and suppressed β-tubulin III (red) expression by IF. B: WB analysis of GFAP and β-tubulin III after rhsAPPα treatment of NSCs. C: Densitometry analysis showed significantly increased ratio of GFAP to β-tubulin III in rhsAPPα treated NSCs compared with heat-inactivated hsAPPα (heat inact.). Data presented as mean ± sd. These results are representative of three independent experiments with triplicates for each condition. *** P < 0.001.

Article Snippet: After blocking with 5% milk in 1X TBS, blots were incubated overnight at 4°C with either of the following primary antibodies: mouse monoclonal anti-Aβ 1-17 (6E10, 1:1,000, Covance Research Products); mouse glial fibrillary acidic protein (GFAP) (1:1,000, Cell Signaling Technology); mouse monoclonal anti-β-tubulin (1:1,000, Stem Cell Technologies, Tukwila, WA); rabbit polyclonal anti-APP C terminus (pAb 396, 1:1,000, kindly provided by S. Gandy and H. Steiner); rabbit polyclonal anti-Notch1 (1:1,000, Epitomics, Burlingame, CA); rabbit polyclonal anti-Notch 1 intercelluar domain (NICD, activated Notch 1) (1:500, Abcam, Cambridge, MA); rabbit polyclonal anti-gp130 (1:1,000, Merck Millipore, Darmstadt, Germany); mouse monoclonal anti-β-actin (1:4,000, Sigma-Aldrich).

Techniques: Recombinant, Expressing

Journal: iScience

Article Title: Sorafenib induces cachexia by impeding transcriptional signaling of the SET1/MLL complex on muscle-specific genes

doi: 10.1016/j.isci.2024.110913

Figure Lengend Snippet: Effect of TKIs on sarcomere organization (A) Panel shows confocal microscopy images of TKI-treated myotubes immunostained against alpha-actinin (green) and nucleus (DAPI – blue). Images were captured at 4× magnification over 60× oil immersion objective. Scale bar = 10 μm. See also Figure S1 . The images are the representative of at least three biological experiments (N). The concentrations used for Nilo was 0.1 μM, and 20 μM for Ima and Sor. 0.1% DMSO was used as vehicle control. (B–D) Western blot showing protein expression of MyHC-IId in myotubes treated with (B) Nilo, (C) Ima, and (D) Sor. (E–G) Western blot showing protein expression of MyHC-IIa in myotubes treated with (E) Nilo, (F) Ima, and (G) Sor. β-tubulin was used as the loading control. The blots are representative of three biological experiments (N). All concentrations are in μM. See also Figure S2 .

Article Snippet: Mouse monoclonal anti-beta tubulin , Biocat , Cat#AKR-009.

Techniques: Confocal Microscopy, Control, Western Blot, Expressing

Journal: iScience

Article Title: Sorafenib induces cachexia by impeding transcriptional signaling of the SET1/MLL complex on muscle-specific genes

doi: 10.1016/j.isci.2024.110913

Figure Lengend Snippet:

Article Snippet: Mouse monoclonal anti-beta tubulin , Biocat , Cat#AKR-009.

Techniques: Recombinant, Isolation, SYBR Green Assay, Protease Inhibitor, Control, Software, Real-time Polymerase Chain Reaction

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