Journal: Journal of Molecular Medicine (Berlin, Germany)

Article Title: Regulation of tamoxifen sensitivity by the PLAC8/MAPK pathway axis is antagonized by curcumin-induced protein stability change

doi: 10.1007/s00109-021-02047-5

Figure Lengend Snippet: Relationship between the ubiquitin of PLAC8 and curcumin. a Western blot (upper) tested the expression level of PLAC8 decreased by curcumin in a dose- and time-dependent manner. RT-PCR (bottom) showed the mRNA expression level of PLAC8 was not changed by curcumin. The expression was quantified and normalized to GAPDH. b Western blot tested the expression level of PLAC8 decreased by treatment of curcumin 24 h (5 μM) could be recused by treatment of MG-132 6 h (10μM) in MCF-7/TAM (upper) and MCF-7 (bottom). c MCF-7/TAM cells were transfected with PLAC8. At 24 h post transfection, cells were treated with or without curcumin (5 μM). After 18 h later, cells were treated with or without MG132 (10 μM) for 6 h and immunoprecipitated with PLAC8 antibody. d Cell proliferation assay showed that curcumin (10 μM) alone or combined with MAPKs inhibitors (SCH772984 10 μM or SB2020190 20 μM) in MCF-7/TAM 24hr. e MCF-7/TAM treated with curcumin (10 μM) alone or combined with MAPKs inhibitors(SCH772984 10 μM or SB2020190 20 μM) and then overexpressing PLAC8 expression. All data represent mean ± SD of three experiments performed in triplicate. ** p < 0.01; * p < 0.05; NS, not significant

Article Snippet: The slices were stained with PLAC8 (1:200) and Ki-67 (1:500) (100130-MM22, Sino Biological, Beijing, China) for 1 h at 37 °C.

Techniques: Western Blot, Expressing, Reverse Transcription Polymerase Chain Reaction, Transfection, Immunoprecipitation, Proliferation Assay

Journal: Journal of Molecular Medicine (Berlin, Germany)

Article Title: Regulation of tamoxifen sensitivity by the PLAC8/MAPK pathway axis is antagonized by curcumin-induced protein stability change

doi: 10.1007/s00109-021-02047-5

Figure Lengend Snippet: Tamoxifen inhibited tumor growth and showed a synergistic activity with curcumin in vivo. ( a ) Body weight ( b ), tumor volume (every 3 days) ( c ) and tumor weight (day 24 after treating drug) ( d ) of xenograft-bearing mice in four groups. 1, Control; 2, Curcumin; 3, Tamoxifen; 4, Curcumin and Tamoxifen. ( ∗* P < 0.01 vs. control group [upper], # P < 0.05 vs. single-treated group). e Western blot analysis results showed that curcumin induced the downregulation of PLAC8 protein levels in MCF-7/TAM xenograft-bearing mice. f IHC staining showed the expression of Ki67 and PLAC8. Scale bar represents 50 μm. g Hypothesis model of curcumin action on regulating BC progression. Our indicated that PLAC8 could induce the tamoxifen resistance, thus affecting the ERK1/2-p38 kinase pathways. In addition, curcumin decreased the level of PLAC8 through ubiquitination modification. This mechanism may explain the potential mechanism of tamoxifen resistance to breast cancer

Article Snippet: The slices were stained with PLAC8 (1:200) and Ki-67 (1:500) (100130-MM22, Sino Biological, Beijing, China) for 1 h at 37 °C.

Techniques: Activity Assay, In Vivo, Western Blot, Immunohistochemistry, Expressing, Modification

Journal: Cell Transplantation

Article Title: Expandable Sendai-Virus-Reprogrammed Human iPSC-Neuronal Precursors: In Vivo Post-Grafting Safety Characterization in Rats and Adult Pig

doi: 10.1177/09636897221107009

Figure Lengend Snippet: Antibodies Used for Immunofluorescence Staining.

Article Snippet: Ab16667 , Ki67 , abcam.

Techniques: Immunofluorescence, Staining

Journal: Cell Transplantation

Article Title: Expandable Sendai-Virus-Reprogrammed Human iPSC-Neuronal Precursors: In Vivo Post-Grafting Safety Characterization in Rats and Adult Pig

doi: 10.1177/09636897221107009

Figure Lengend Snippet: Similar differentiation profile of induced-proliferating NPCs or previously frozen and induced NPCs in vitro . (A, B) Bright field morphology of proliferating and previously frozen-washed and plated NPCs. The image from previously frozen cells was taken at 48 h after plating in NPC media. (C, D) Co-staining with Nestin and MAP2 antibodies shows a mixed population of Nestin+, MAP2+, or Nestin/MAP2 expressing cells. (E, F) Presence of early postmitotic DCX+ neurons and early glial VIM + precursors. (G, H) Staining with the Tuj1 antibody show well-developed neurons with axodendritic arborization. In the same field, a high density of mitotically active cells (Ki67+) is also seen. (I) Quantitative analysis of Nestin, MAP2, DCX, Tuj1, and Ki67-positive cells showed no significant difference between freshly induced and frozen-washed and induced NPCs. Scale bars: 200 µm (A–H). NPC: neural precursors cell; VIM: Vimentin; DAPI: 4′,6-diamidino-2-phenylindole.

Article Snippet: Ab16667 , Ki67 , abcam.

Techniques: In Vitro, Staining, Expressing

Journal: Cell Transplantation

Article Title: Expandable Sendai-Virus-Reprogrammed Human iPSC-Neuronal Precursors: In Vivo Post-Grafting Safety Characterization in Rats and Adult Pig

doi: 10.1177/09636897221107009

Figure Lengend Snippet: Comparable survival and differentiation of freshly harvested or previously frozen-washed NPCs grafted into striata of the immunodeficient rat. (A, B) Comparable engraftment of hNUMA + cells with DCX and hGFAP immunoreactivity in the core of the graft. (C, D) Expression of NeuN in hNUMA+ neurons. Homogenous vimentin IF in the same areas can also be seen. (E, F) Sparse presence of grafted cells-derived oligodendrocytes (Olig2). (G, H) Co-expression of Ki67 mitotic marker in grafted hNUMA cells. (I) Quantitative analysis of neuronal, glial, and mitotic marker expression in grafted cells. No significant differences in differentiation profile between NPCs grafted as freshly harvested or previously frozen cells can be seen. Scale bars: 20 µm (A-H), 10 µm (A-H inserts). NPC: neural precursors cell; VIM: Vimentin; DAPI: 4′,6-diamidino-2-phenylindole; hGFAP-human-specific glial fibrillary acidic protein.

Article Snippet: Ab16667 , Ki67 , abcam.

Techniques: Expressing, Derivative Assay, Marker

Journal: Cell Transplantation

Article Title: Expandable Sendai-Virus-Reprogrammed Human iPSC-Neuronal Precursors: In Vivo Post-Grafting Safety Characterization in Rats and Adult Pig

doi: 10.1177/09636897221107009

Figure Lengend Snippet: Survival and differentiation of freshly harvested or previously frozen-washed NPCs grafted into the lumbar spinal cord of the immunodeficient rat. (A, B) Presence of hNUMA+ cells with dense DCX immunoreactivity in early postmitotic neurons. hGFAP immunoreactivity throughout the grafts can also be seen. (C, D) NeuN expression in mature neurons in the core of hNUMA+ grafts. Vimentin positivity was typically seen at the periphery of the grafts. (E, F) Presence of grafted cells-derived oligodendrocytes (Olig2) throughout the grafts. (G, H) The occasional presence of mitotically active cells (Ki67) in hNUMA + grafts. (I) Quantitative analysis of neuronal, glial, and mitotic marker expression in grafted cells. No significant differences between NPCs grafted as freshly -harvested or previously frozen cells were identified. Scale bars: 20 µm (A-H), 10 µm (A-H inserts). NPC: neural precursors cell; VIM: Vimentin; DAPI: 4′,6-diamidino-2-phenylindole; hGFAP-human-specific glial fibrillary acidic protein.

Article Snippet: Ab16667 , Ki67 , abcam.

Techniques: Expressing, Derivative Assay, Marker

Journal: Frontiers in Pediatrics

Article Title: Neurogenesis Is Reduced at 48 h in the Subventricular Zone Independent of Cell Death in a Piglet Model of Perinatal Hypoxia-Ischemia

doi: 10.3389/fped.2022.793189

Figure Lengend Snippet: Fluorescent microphotographs of Ki67 [cell proliferation, (A,B) ], Sox2/Ki67 [neural stem/progenitor cell proliferation, (C,D) ], and DAPI [nuclei, (E,F) ] immune-stained samples from control and HI piglets. Ki67 positive cells (green) are abundant in the control piglet (A) , whereas HI (B) significantly reduced its counts by half in L-SVZ and DL-SVZ (both *** p < 0.001). Double Sox2 (red) + /Ki67 (green) + cells are observed in control animals; again, HI reduced neural stem/progenitor cell counts in L-SVZ and DL-SVZ (both *** p < 0.001). DAPI immunostaining reveals total nuclei. LV, lateral ventricle; SVZ, subventricular zone; CDT, caudate nucleus; PvWM, periventricular white matter. Original magnification 400×. Scale bar: 100 μm.

Article Snippet: For single labeling, radial-glia/neural stem cells were identified using an anti-glial fibrillary acidic protein (GFAP) antibody to assess neurogenic activity close to the ventricular wall (mouse anti-GFAP, 1:100, MA5-12023, Thermo Fisher, United States); an anti-doublecortin (DCX) antibody was used to identify young neurons/neuroblasts (mouse anti-DCX, 1:50, sc-271390, Santa Cruz Biotechnology, United States); cell proliferation was identified by an anti-Ki67 antibody (mouse anti-Ki67; 1:50, STJ96966, St Johns Labs, United Kingdom).

Techniques: Staining, Control, Immunostaining

Journal: Oncology letters

Article Title: Solamargine triggers hepatoma cell death through apoptosis.

doi: 10.3892/ol.2015.3194

Figure Lengend Snippet: Figure 6. (A) Expression of apoptosis‑associated proteins in SMMC7721 and HepG2 cells treated with SM and (B) the statistical results of this analysis. The expression levels of Bcl‑2, Bax, caspase‑3 and caspase‑9 were detected by western blot analysis using β‑actin as a control. (C) Expression levels of proliferation‑associated proteins, ki67 and pcna, also investigated by western blot analysis and (D) the statistical results of this analysis. The results are representative of at least three independent experiments. SM, solamargine.

Article Snippet: Rabbit anti-human polyclonal Ki67 primary antibody (1:500; cat. no. BA1508) was purchased from Wuhan Boster Biotechnology, Ltd., (Wuhan, China).

Techniques: Expressing, Western Blot, Control

Journal: Cells

Article Title: Cistanoside F Ameliorates Lipid Accumulation and Enhances Myogenic Differentiation via AMPK-Dependent Signaling in C2C12 Myotubes.

doi: 10.3390/cells14120874

Figure Lengend Snippet: Figure 4. Expression of PGC-1α, MMP, and ROS in two adipogenic models after Cis adminis- tration. (A,B) PGC-1α expression. (C,D) Immunofluorescence of TMRE, the statistic was eval- uated by fluorescence spectroscopy, at λexcitation = 550 nm and λemission = 575 nm. (E,G) Im- munofluorescence of JC-1, statistic was evaluated by fluorescence spectroscopy, JC-1 monomer at λexcitation = 490 nm and λemission = 530 nm and JC-1 aggregate at λexcitation = 525 nm and λemission = 590 nm. (F,H) Immunofluorescence of ROS, the statistic was evaluated by fluorescence spectroscopy, at λexcitation = 488 nm and λemission = 525 nm. n = 6. ## p < 0.01, compared with the vehicle group; * p < 0.05, ** p < 0.01, compared with the model group.

Article Snippet: Antibodies against UCP1, CPT1b, ACC2, Desmin, and PGC-1α were purchased from Boster Biotech (Wuhan, China; #M00255; #A03558; #A03668; #PB9105; #BM4898; 1:1000).

Techniques: Expressing, Immunofluorescence, Fluorescence, Spectroscopy