Journal: Experimental and Therapeutic Medicine

Article Title: Harmine reinforces the effects of regorafenib on suppressing cell proliferation and inducing apoptosis in liver cancer cells

doi: 10.3892/etm.2022.11132

Figure Lengend Snippet: Harmine combined with regorafenib induces cell death via the AKT pathway. (A) Liver cancer cells were incubated with harmine/regorafenib for 48 h and western blot analysis was performed to detect protein expression. (B) The expression of indicated proteins was determined. One-way ANOVA followed by Tukey's post hoc test was used to examine the significant differences among four groups, including control, regorafenib, harmine and regorafenib plus harmine groups. ** P<0.01 and **** P<0.0001. ns, no significance; Rego, regorafenib; Har, harmine; Mcl-1, myeloid cell leukemia-1; PARP, poly (ADP-ribose) polymerase; p-, phosphorylated.

Article Snippet: The primary antibodies against PARP-1/2 (H-250) (cat. no. sc-7150; 1:500), anti-GAPDH antibody (FL-335) (cat. no. sc-25778; 1:500), myeloid cell leukemia-1 (Mcl-1) (S-19) (cat. no. sc-819; 1:500), phosphorylated (p)-AKT (1/2/3) (Ser473) (cat. no. sc-7985; 1:500) and caspase-3 (H-277) (cat. no. sc-7148; 1:500) were purchased from Santa Cruz Biotechnology, Inc.

Techniques: Incubation, Western Blot, Expressing, Control

Journal: Experimental and Therapeutic Medicine

Article Title: Harmine reinforces the effects of regorafenib on suppressing cell proliferation and inducing apoptosis in liver cancer cells

doi: 10.3892/etm.2022.11132

Figure Lengend Snippet: Effect and mechanism of harmine plus regorafenib. DYRK1A, dual-specificity tyrosine phosphorylation-regulated kinase 1A; Mcl-1, myeloid cell leukemia-1; si-, small interfering.

Article Snippet: The primary antibodies against PARP-1/2 (H-250) (cat. no. sc-7150; 1:500), anti-GAPDH antibody (FL-335) (cat. no. sc-25778; 1:500), myeloid cell leukemia-1 (Mcl-1) (S-19) (cat. no. sc-819; 1:500), phosphorylated (p)-AKT (1/2/3) (Ser473) (cat. no. sc-7985; 1:500) and caspase-3 (H-277) (cat. no. sc-7148; 1:500) were purchased from Santa Cruz Biotechnology, Inc.

Techniques: Phospho-proteomics

Journal: Journal of Biological Chemistry

Article Title: RNA Polymerase II C-terminal Heptarepeat Domain Ser-7 Phosphorylation Is Established in a Mediator-dependent Fashion

doi: 10.1074/jbc.m109.046565

Figure Lengend Snippet: FIGURE 3. CDK7 is a CTD Ser-7 kinase. A, shown is an immobilized template assay on the pGL2-MRG5 pro- moter template in the presence of the activator GAL-VP16. Mock-treated or CDK7-, CDK8-, and CDK9-depleted nuclear extracts (lanes 5–8) were used in the PIC formation reactions. Reactions were carried out under stand- ard in vitro transcription conditions in the presence of 1 M ATP. Lanes 1–4 show 20% of reaction input. B, shown is an immobilized template assay on the ML-promoter template in the presence of GAL-VP16 using mock-treated (Iso), core-TFIIH (p62), CDK7 (CDK7)- or CDK8-depleted (CDK8) Jurkat nuclear extract. Lanes 1–4 show 20% reaction input. TXN, in vitro transcription analysis. C, shown is an immobilized template assay on ML-promoter DNA templates under basal conditions at physiological salt concentrations. PICs were formed in the presence of the indicated amounts of ATPS, washed, and then analyzed by immunoblot. D, shown is an immobilized template assay on the ML-promoter DNA template using mock-treated (Iso), Mediator-depleted (MED), and CDK7-depleted (CDK7) 0.5 M P11 fractions together with GAL4-VP16, recombinant yeast TBP, Toa, and recombinant human TFIIB as indicated. In lane 6 a high salt-washed MED15-IP was added. PIC forma- tion was carried out under physiological conditions in the presence of 1 M ATP and 10 M ATPS. NEX, nuclear extract.E,immobilizedtemplateassayonML-promoterDNAtemplatesusingeitherJurkatnuclearextract(lane 1) or a recombinant transcription system consisting of yeast TBP, Toa, human TFIIB, TFIIE, and TFIIF) in combi- nation with a high salt-washed Mediator-IP (which also provides RNAPII; lanes 2 and 3) is shown. In lane 3 a stringently washed CDK7 IP was also added to the PIC formation reactions. PIC formation in D and E was carried out under physiological conditions in the presence of 1 M ATP and 10 M ATPS.

Article Snippet: Jurkat nuclear extracts were immunodepleted using antiRNAPII (1C7), MED15 (1H7), and MED25 (VC1) rat monoclonal antibodies, anti-TBP, TFIIH p62, CDK7, and CDK9 rabbit polyclonal antibodies (Santa Cruz sc-273, sc-292, sc- 529, and sc-8338, respectively), and anti-TFIIB (IIB8) mouse monoclonal antibodies (Santa Cruz sc-23875) or anti-CDK8 goat polyclonal antibodies (Santa Cruz sc-1521).

Techniques: In Vitro, Western Blot, Recombinant