Journal: Scientific Reports

Article Title: Regulation of cardiomyocyte DNA damage and cell death by the type 2A protein phosphatase regulatory protein alpha4

doi: 10.1038/s41598-021-85616-5

Figure Lengend Snippet: Regulation of the DNA damage response by α4 protein expression in cardiomyocytes. H9c2 cardiomyocytes were transfected with either 50 nM non-targeting control (NTC) siRNA or 50 nM siRNA specific to rat α4 protein (siα4) for 1–4 days (n = 4) and the KD of α4 protein expression was confirmed by Western analysis using an antibody raised against human α4 ( a ). α4 protein expression was knocked down for 4 days and Western analysis (short and long exposures) was used to probe samples (n = 3) with an antibody specific to phosphorylated S / T QG motifs within ATM/ATR substrates. Arrows indicate substrates whose phosphorylation status (n = 3) was elevated in response to α4 KD ( b ). The expression and phosphorylation (Ser15) of p53 were determined by Western analysis using rodent-specific antibodies raised against mouse p53 protein in samples following 4 days of α4 KD, n = 5 ( c ). The expression of PARP was determined by Western analysis (short and long exposures) following 1–4 days of α4 KD, n = 3. Arrows indicate full length PARP (116 kDa) and cleaved fragments (89 kDa and 24 kDa) ( d ). All data represents data mean values ± SEM and statistical significance vs. non-targeting control (NTC) was determined by a one-tailed Student’s T-test. Expression of β-actin was used to confirm equal protein loading between samples.

Article Snippet: Antibodies to detect Bax (#2772), Bcl-xL (#2762), phosphospecific ATM/ATR substrate (#6966), IGBP1 (α4) (#5699), PARP (#9542), phosphospecific (Ser139) H2AX (#9718), H2AX (#2595), rodent-specific p53 (#32532), rodent-specific phosphospecific (Ser15) p53 (#12571) and V5-tag (#13202) were obtained from Cell Signaling Technology (USA).

Techniques: Expressing, Transfection, Control, Western Blot, Phospho-proteomics, One-tailed Test

Journal: Cell reports

Article Title: p53 Promotes Cancer Cell Adaptation to Glutamine Deprivation by Upregulating Slc7a3 to Increase Arginine Uptake

doi: 10.1016/j.celrep.2019.02.037

Figure Lengend Snippet: (A) Immunoblot for phospho-p53 (S18), total p53, and actin in MEF cells cultured in complete or glutamine-free medium for 18 h.

Article Snippet: Immunoblotting was performed with the following antibodies: p53, rodent (CM5) was purchased from VECTOR Laboratories, Burlingame, CA, USA; p53, human (DO-1, SC126) and CAT-3, mouse (T-14, sc161433) were purchased from Santa Cruz Biotechnology, Santa Cruz, CA, USA; phospho-p53 (Ser15, #9284), p53 rodent specific (D2H90, #32532), phospho-p70S6K (Thr389, #9234), p70S6K (#2708) and ATF4 (D4B8, #11815) were purchased from Cell Signaling Technology, Danvers, MA, USA, and beta actin (A1978) from Sigma-Aldrich, St. Louis, MO, USA.

Techniques: Western Blot, Cell Culture

Journal: Cell reports

Article Title: p53 Promotes Cancer Cell Adaptation to Glutamine Deprivation by Upregulating Slc7a3 to Increase Arginine Uptake

doi: 10.1016/j.celrep.2019.02.037

Figure Lengend Snippet: (A) Relative arginine levels of MEF WT and p53−/− cells cultured in complete or glutamine-free medium for 6 h.

Article Snippet: Immunoblotting was performed with the following antibodies: p53, rodent (CM5) was purchased from VECTOR Laboratories, Burlingame, CA, USA; p53, human (DO-1, SC126) and CAT-3, mouse (T-14, sc161433) were purchased from Santa Cruz Biotechnology, Santa Cruz, CA, USA; phospho-p53 (Ser15, #9284), p53 rodent specific (D2H90, #32532), phospho-p70S6K (Thr389, #9234), p70S6K (#2708) and ATF4 (D4B8, #11815) were purchased from Cell Signaling Technology, Danvers, MA, USA, and beta actin (A1978) from Sigma-Aldrich, St. Louis, MO, USA.

Techniques: Cell Culture

Journal: Cell reports

Article Title: p53 Promotes Cancer Cell Adaptation to Glutamine Deprivation by Upregulating Slc7a3 to Increase Arginine Uptake

doi: 10.1016/j.celrep.2019.02.037

Figure Lengend Snippet: (A) Immunoblot of phospho-S6K (T389), total S6K, total p53, and actin in SaOs-2 cells cultured in the absence or presence of doxycycline for 24 h and then separated into complete and glutamine-free medium for 3-h increments.

Article Snippet: Immunoblotting was performed with the following antibodies: p53, rodent (CM5) was purchased from VECTOR Laboratories, Burlingame, CA, USA; p53, human (DO-1, SC126) and CAT-3, mouse (T-14, sc161433) were purchased from Santa Cruz Biotechnology, Santa Cruz, CA, USA; phospho-p53 (Ser15, #9284), p53 rodent specific (D2H90, #32532), phospho-p70S6K (Thr389, #9234), p70S6K (#2708) and ATF4 (D4B8, #11815) were purchased from Cell Signaling Technology, Danvers, MA, USA, and beta actin (A1978) from Sigma-Aldrich, St. Louis, MO, USA.

Techniques: Western Blot, Cell Culture

Journal: Cell reports

Article Title: p53 Promotes Cancer Cell Adaptation to Glutamine Deprivation by Upregulating Slc7a3 to Increase Arginine Uptake

doi: 10.1016/j.celrep.2019.02.037

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Immunoblotting was performed with the following antibodies: p53, rodent (CM5) was purchased from VECTOR Laboratories, Burlingame, CA, USA; p53, human (DO-1, SC126) and CAT-3, mouse (T-14, sc161433) were purchased from Santa Cruz Biotechnology, Santa Cruz, CA, USA; phospho-p53 (Ser15, #9284), p53 rodent specific (D2H90, #32532), phospho-p70S6K (Thr389, #9234), p70S6K (#2708) and ATF4 (D4B8, #11815) were purchased from Cell Signaling Technology, Danvers, MA, USA, and beta actin (A1978) from Sigma-Aldrich, St. Louis, MO, USA.

Techniques: Plasmid Preparation, Control, Virus, Recombinant, Protease Inhibitor, cDNA Synthesis, SYBR Green Assay, Transformation Assay, Generated, shRNA

Journal: Asian Pacific Journal of Tropical Biomedicine

Article Title: Hepatorenoprotective effects of Lepidium draba L. extracts against cyclophosphamide-induced oxidative injuries in rats via reducing apoptosis and inflammation

doi: 10.4103/apjtb.apjtb_518_24

Figure Lengend Snippet: Figure 6. Effects of L. draba extract on p53-positive cells in (A) kidney and liver tissues (DAB staining 400×, scale bar = 25 μm). (B) Quantitative results of immunohistochemical examination. Data are shown as mean ± SEM (n=6).

Article Snippet: The slides were then incubated in p53 rodent-specific monoclonal antibody (Cat. no. sc-98, Pab 1801, dilution 1꞉200, Santa Cruz Biotechnology) for 45 min at room temperature, then incubated for 5 min in 5% bovine serum albumin to block unbound antibodies, and finally in chromogen (3, 3’-diaminobenzidine) for 30 min at 37 曟. Counterstaining of the slides was performed with hematoxylin and 50 fields of view were randomly evaluated with a light microscope (Olympus CH3, Japan) attached to the camera (Moticam Technologies, Japan), and the p53 positive cells/total cells × 100 was calculated at 400× magnification and reported as mean (%) ± SEM[23].

Techniques: Staining, Immunohistochemical staining