anti-pp2ac antibody Search Results


metyl-pp2ac  (Merck & Co)
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Merck & Co metyl-pp2ac
A Western blot experiment showing a time-course analysis of LC3-II/LC3-I. PANC1 cells were treated with vehicle or HMT for 48 h followed by treatment with CQ (40 µM) for the indicated times. * P < 0.05. ns = not significant. Differences between vehicle and HMT-treated samples at each time were found significant ( # P < 0.05). B Pancreatic cancer cells were treated with vehicle or HMT for 48 h followed (or not) by treatment with CQ (40 µM) for an additional 24 h. Accumulated autophagosomes were evaluated by LC3 staining. * P < 0.05 when compared with vehicle-treated cells without CQ (w/o). C Accumulation of p62 was analyzed by confocal microscopy (left) and Western blot (right) in PANC1 cells after 72 h of treatment with vehicle or HMT. D Effect of HMT and individual treatments [TMCG (10 µM) or DIPY (5 µM)] on the methylation capacity of PANC1 cells. Metabolite determination was performed 72 h after treatment. E Effect of HMT and AdOx (20 µM) on the methylation status (at L309) of the catalytic <t>PP2Ac</t> subunit. The histogram shows the effect of HMT on PP2A activity. Protein and activity analysis was carried out 72 h after each indicated treatment. In this figure, blots are representative of three independent experiments. Error bars show the mean ± SD. * P < 0.05 compared with vehicle-treated controls.
Metyl Pp2ac, supplied by Merck & Co, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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metyl-pp2ac - by Bioz Stars, 2026-11
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A Western blot experiment showing a time-course analysis of LC3-II/LC3-I. PANC1 cells were treated with vehicle or HMT for 48 h followed by treatment with CQ (40 µM) for the indicated times. * P < 0.05. ns = not significant. Differences between vehicle and HMT-treated samples at each time were found significant ( # P < 0.05). B Pancreatic cancer cells were treated with vehicle or HMT for 48 h followed (or not) by treatment with CQ (40 µM) for an additional 24 h. Accumulated autophagosomes were evaluated by LC3 staining. * P < 0.05 when compared with vehicle-treated cells without CQ (w/o). C Accumulation of p62 was analyzed by confocal microscopy (left) and Western blot (right) in PANC1 cells after 72 h of treatment with vehicle or HMT. D Effect of HMT and individual treatments [TMCG (10 µM) or DIPY (5 µM)] on the methylation capacity of PANC1 cells. Metabolite determination was performed 72 h after treatment. E Effect of HMT and AdOx (20 µM) on the methylation status (at L309) of the catalytic PP2Ac subunit. The histogram shows the effect of HMT on PP2A activity. Protein and activity analysis was carried out 72 h after each indicated treatment. In this figure, blots are representative of three independent experiments. Error bars show the mean ± SD. * P < 0.05 compared with vehicle-treated controls.

Journal: Cell Death & Disease

Article Title: Targeting protein methylation in pancreatic cancer cells results in KRAS signaling imbalance and inhibition of autophagy

doi: 10.1038/s41419-023-06288-9

Figure Lengend Snippet: A Western blot experiment showing a time-course analysis of LC3-II/LC3-I. PANC1 cells were treated with vehicle or HMT for 48 h followed by treatment with CQ (40 µM) for the indicated times. * P < 0.05. ns = not significant. Differences between vehicle and HMT-treated samples at each time were found significant ( # P < 0.05). B Pancreatic cancer cells were treated with vehicle or HMT for 48 h followed (or not) by treatment with CQ (40 µM) for an additional 24 h. Accumulated autophagosomes were evaluated by LC3 staining. * P < 0.05 when compared with vehicle-treated cells without CQ (w/o). C Accumulation of p62 was analyzed by confocal microscopy (left) and Western blot (right) in PANC1 cells after 72 h of treatment with vehicle or HMT. D Effect of HMT and individual treatments [TMCG (10 µM) or DIPY (5 µM)] on the methylation capacity of PANC1 cells. Metabolite determination was performed 72 h after treatment. E Effect of HMT and AdOx (20 µM) on the methylation status (at L309) of the catalytic PP2Ac subunit. The histogram shows the effect of HMT on PP2A activity. Protein and activity analysis was carried out 72 h after each indicated treatment. In this figure, blots are representative of three independent experiments. Error bars show the mean ± SD. * P < 0.05 compared with vehicle-treated controls.

Article Snippet: Antibodies against the following proteins were used: β-actin (Merck; monoclonal clone AC-15), AKT (Abcam, Cambridge, UK; monoclonal clone EPR17737), ATF4 (Cell Signaling Technology, Danvers, MA, USA; polyclonal H-277), Beclin-1 (Thermo-Fisher; monoclonal), Bcl-2 (Merck; monoclonal), BRAF (Abcam; monoclonal clone EP152Y), caspase 3 (Santa Cruz Biotechnology, Dallas, TX, USA; monoclonal), c-Jun (Thermo-Fisher; recombinant polyclonal clone 2HCLC), CRAF (Thermo-Fisher; polyclonal), EGFR (Abcam; monoclonal clone EP38Y), ERK1/2 (Santa Cruz Biotechnology; monoclonal), GADPH (Abcam; monoclonal), Lamp-1 (Abcam; polyclonal), LC3B (Abcam, polyclonal), MEK1/2 (Thermo-Fisher; polyclonal), metyl-PP2Ac (Merck; monoclonal clone 2A10), MITF (Merck; monoclonal clone C5), MRAS (Thermo-Fisher; monoclonal clone OTI3C4), MRAS (Abcam; monoclonal clone EPR12457), p62/SQSTM1 (Thermo-Fisher; polyclonal), p-AKT-S473 (Merck; monoclonal clone 6F5), p-AKT-T308 (Santa Cruz Biotechnology; monoclonal), PARP (Abcam; monoclonal clone E102), p-Bcl-2-S70 (Merck; polyclonal), p-c-Jun (Thermo-Fisher; polyclonal), p-CRAF-S259 (Thermo-Fisher; polyclonal), p-eIF2α (Cell Signaling Technology; monoclonal), p-ERK1/2 (Merck; monoclonal), PGC-1α (Santa Cruz Biotechnology; monoclonal), p-MEK1/2-S217-S221 (Thermo-Fisher; monoclonal clone C.158.9), PP2Ac (Merck; monoclonal clone 1D6), p-S6K-T389 (Merck; polyclonal B2HCLC), p-ULK1-S638 (Abcam; monoclonal clone EPR6155), S6K (Merck; monoclonal clone 5G9), SHOC2 (Merck; polyclonal), TATA binding protein (TBP: Abcam; monoclonal), TFE3 (Merck; polyclonal), TFEB (Merck; polyclonal), and ULK1 (Merck; polyclonal).

Techniques: Western Blot, Staining, Confocal Microscopy, Methylation, Activity Assay

A In normal cells and under nutrient-rich conditions, MiT/TFE proteins are repressed by mTORC1 via phosphorylation and remain in the cytosol; however, upon starvation, mTORC1 is inactivated and MiT/TFE proteins enter the nucleus, where they activate autophagy-lysosome gene transcription. Interestingly, this mechanism was found to be decoupling in PDAC . Although overexpression of the nucleocytoplasmic transporter importin (IPO-7/8) was proposed to facilitate the nuclear translocation of MiT/TFE, the high PP2A activity found in autophagic PDAC cells could also explain the MiT/TFE factors decoupling mechanism. The possible mechanism by which HMT inhibits basal autophagy in PDCA is also depicted. B Efficient PP2Ac silencing in PANC1 cells promotes cytosolic accumulation of MITF and TFE3 factors (* P < 0.05 when compared with siControl transfected cells). Blots were overexposed (TFE3 over ) for cytosolic TFE3 quantification. C Fluorescence microscopy showing the effect of PP2Ac silencing on the localization of TFE3 in Hs766T cells. The line bar (30 µm) was used as a template for fluorescence intensity determination. Relative occupancy of TFE3 in the nucleus (Nu) and cytosol (Cyt) was calculated by analysis of the fluorescence intensity under the curve (IUC) in each compartment. * P < 0.05 when compared with siControl transfected cells. D HMT promotes cytosolic accumulation of MITF and TFE3 factors in PANC1 cells (* P < 0.05 when compared with vehicle-treated cells). Blots were overexposed (TFE3 over ) for cytosolic TFE3 quantification. E HMT avoids nuclear translocation of TFE3 and MITF in Hs766T cells. Cells were treated with vehicle or HMT (72 h) or rapamycin (0.2 µM; 120 min). Cell extracts were analyzed by Western blotting for the indicated proteins (left panel). Nuclear localization of MITF and TFE3 was also evaluated by confocal microscopy (right panels) in Hs766T cells treated with vehicle or HMT (72 h). F Relative expression of proteins 72 h after each indicated treatment. Histogram represents the relative amount of proteins with respect to actin. * P < 0.05 when compared with vehicle-treated cells. G HMT treatment (72 h) causes aberrant lysosomal morphology and increased size as shown by LysoTracker staining. Inset: magnified view. Graph (right) quantification of lysosome diameter in HMT ( N = 173) and vehicle-treated cells ( N = 169). Bar, mean. H Confocal microscopy showing lysosome number and distribution in autophagic cells and HMT-treated cells. Lysosomes were stained with LysoTracker-Red DND-99 and an iABP Pan cathepsin probe (green). Differences in the number of lysosomes were significant (* P < 0.01) when compared with untreated autophagic cells (graph, right). I TEM showing lysosome localization in autophagic cells and HMT-treated cells. The arrow indicates the position of lysosomes. The boxed area contains representative lysosomes at 60,000× magnification. Lysosomes were randomly distributed in the cytoplasm of autophagic PANC1 cells, but HMT-treated cells showed grouped lysosomes.

Journal: Cell Death & Disease

Article Title: Targeting protein methylation in pancreatic cancer cells results in KRAS signaling imbalance and inhibition of autophagy

doi: 10.1038/s41419-023-06288-9

Figure Lengend Snippet: A In normal cells and under nutrient-rich conditions, MiT/TFE proteins are repressed by mTORC1 via phosphorylation and remain in the cytosol; however, upon starvation, mTORC1 is inactivated and MiT/TFE proteins enter the nucleus, where they activate autophagy-lysosome gene transcription. Interestingly, this mechanism was found to be decoupling in PDAC . Although overexpression of the nucleocytoplasmic transporter importin (IPO-7/8) was proposed to facilitate the nuclear translocation of MiT/TFE, the high PP2A activity found in autophagic PDAC cells could also explain the MiT/TFE factors decoupling mechanism. The possible mechanism by which HMT inhibits basal autophagy in PDCA is also depicted. B Efficient PP2Ac silencing in PANC1 cells promotes cytosolic accumulation of MITF and TFE3 factors (* P < 0.05 when compared with siControl transfected cells). Blots were overexposed (TFE3 over ) for cytosolic TFE3 quantification. C Fluorescence microscopy showing the effect of PP2Ac silencing on the localization of TFE3 in Hs766T cells. The line bar (30 µm) was used as a template for fluorescence intensity determination. Relative occupancy of TFE3 in the nucleus (Nu) and cytosol (Cyt) was calculated by analysis of the fluorescence intensity under the curve (IUC) in each compartment. * P < 0.05 when compared with siControl transfected cells. D HMT promotes cytosolic accumulation of MITF and TFE3 factors in PANC1 cells (* P < 0.05 when compared with vehicle-treated cells). Blots were overexposed (TFE3 over ) for cytosolic TFE3 quantification. E HMT avoids nuclear translocation of TFE3 and MITF in Hs766T cells. Cells were treated with vehicle or HMT (72 h) or rapamycin (0.2 µM; 120 min). Cell extracts were analyzed by Western blotting for the indicated proteins (left panel). Nuclear localization of MITF and TFE3 was also evaluated by confocal microscopy (right panels) in Hs766T cells treated with vehicle or HMT (72 h). F Relative expression of proteins 72 h after each indicated treatment. Histogram represents the relative amount of proteins with respect to actin. * P < 0.05 when compared with vehicle-treated cells. G HMT treatment (72 h) causes aberrant lysosomal morphology and increased size as shown by LysoTracker staining. Inset: magnified view. Graph (right) quantification of lysosome diameter in HMT ( N = 173) and vehicle-treated cells ( N = 169). Bar, mean. H Confocal microscopy showing lysosome number and distribution in autophagic cells and HMT-treated cells. Lysosomes were stained with LysoTracker-Red DND-99 and an iABP Pan cathepsin probe (green). Differences in the number of lysosomes were significant (* P < 0.01) when compared with untreated autophagic cells (graph, right). I TEM showing lysosome localization in autophagic cells and HMT-treated cells. The arrow indicates the position of lysosomes. The boxed area contains representative lysosomes at 60,000× magnification. Lysosomes were randomly distributed in the cytoplasm of autophagic PANC1 cells, but HMT-treated cells showed grouped lysosomes.

Article Snippet: Antibodies against the following proteins were used: β-actin (Merck; monoclonal clone AC-15), AKT (Abcam, Cambridge, UK; monoclonal clone EPR17737), ATF4 (Cell Signaling Technology, Danvers, MA, USA; polyclonal H-277), Beclin-1 (Thermo-Fisher; monoclonal), Bcl-2 (Merck; monoclonal), BRAF (Abcam; monoclonal clone EP152Y), caspase 3 (Santa Cruz Biotechnology, Dallas, TX, USA; monoclonal), c-Jun (Thermo-Fisher; recombinant polyclonal clone 2HCLC), CRAF (Thermo-Fisher; polyclonal), EGFR (Abcam; monoclonal clone EP38Y), ERK1/2 (Santa Cruz Biotechnology; monoclonal), GADPH (Abcam; monoclonal), Lamp-1 (Abcam; polyclonal), LC3B (Abcam, polyclonal), MEK1/2 (Thermo-Fisher; polyclonal), metyl-PP2Ac (Merck; monoclonal clone 2A10), MITF (Merck; monoclonal clone C5), MRAS (Thermo-Fisher; monoclonal clone OTI3C4), MRAS (Abcam; monoclonal clone EPR12457), p62/SQSTM1 (Thermo-Fisher; polyclonal), p-AKT-S473 (Merck; monoclonal clone 6F5), p-AKT-T308 (Santa Cruz Biotechnology; monoclonal), PARP (Abcam; monoclonal clone E102), p-Bcl-2-S70 (Merck; polyclonal), p-c-Jun (Thermo-Fisher; polyclonal), p-CRAF-S259 (Thermo-Fisher; polyclonal), p-eIF2α (Cell Signaling Technology; monoclonal), p-ERK1/2 (Merck; monoclonal), PGC-1α (Santa Cruz Biotechnology; monoclonal), p-MEK1/2-S217-S221 (Thermo-Fisher; monoclonal clone C.158.9), PP2Ac (Merck; monoclonal clone 1D6), p-S6K-T389 (Merck; polyclonal B2HCLC), p-ULK1-S638 (Abcam; monoclonal clone EPR6155), S6K (Merck; monoclonal clone 5G9), SHOC2 (Merck; polyclonal), TATA binding protein (TBP: Abcam; monoclonal), TFE3 (Merck; polyclonal), TFEB (Merck; polyclonal), and ULK1 (Merck; polyclonal).

Techniques: Over Expression, Translocation Assay, Activity Assay, Transfection, Fluorescence, Microscopy, Western Blot, Confocal Microscopy, Expressing, Staining