Table 1.
Compounds tested as inhibitors of G9R‐p27 phosphorylation of serine 12. The compounds reported in the Table were added at the showed concentrations to PC‐3 cultures 2 h before pcDNA3.0 G9R‐p27 or S10A/G9R‐p27 transfection. After 24 h of transfection, PC‐3 cellular extracts were prepared and analyzed by 2D/WB. The bidimensional patterns were analyzed by imagej software to estimate the percentage of inhibition. The values are the mean ± SD of, at least, three separate experiments. S10A/G9R‐p27 transfection was used to evaluate only S12 G9R‐p27 phosphorylation. ND, not done.
| Compounds | Target | Conc (µm) | Inhibitory effect on G9R‐p27 phosphorylation (%) |
Inhibitory effect on S10A/G9R‐p27 Phosphorylation (%) |
|---|---|---|---|---|
| PD‐98059 | MEK | 10 | 0 | 20 ± 5 |
| U‐0126 | MEK | 10 | 0 | 0 |
| SB‐203580 |
P38 MAP Kinase |
10 | 40 ± 5 | ND |
| H‐7‐2HCl | PKA, PKG, MLCK and PKC | 5 | 0 | 0 |
| H‐9‐2HCl | PKA, PKG, MLCK and PKC | 5 | 0 | 0 |
| Staurosporine | Pan‐specific | 0.02 | 0 | 0 |
| Tyrphostin 46 | EGFRK, PDGFRK | 50 | 0 | 0 |
| PKC‐412 | PKC Inhibitor | 5 | 0 | 0 |
| AG‐490 | JAK‐2 | 5 | 0 | 0 |
| LY 294002 | PI 3‐K | 10 | 0 | 0 |
| Wortmannin | PI 3‐K | 5 | 0 | 0 |
| GF 109203X | PKC | 5 | 0 | 0 |
| Hypercin | PKC | 10 | 0 | 0 |
| Ro 31‐8229 mesylate | PKC | 5 | ND | 40 ± 10 |
| D‐erythro‐sphingosine | PKC and CaMK | 5 | 0 | 0 |
| H‐89‐2HCl | PKA | 1 | 0 | 0 |
| 2‐Hydroxy‐5‐(2,5‐dihydroxybenzylamino) benzoic acid | PKA | 5 | ND | 0 |
| KN‐62 | EGFRK, CaMK II | 1 | 60 ± 10 | 70 ± 10 |
| KN‐93 | CaMK II | 1 | ND | 40 ± 10 |
| ML‐9‐HCl | MLCK | 10 | ND | 0 |
| N9‐Isopropyl‐olomucine | CDK | 2 | ND | 0 |
| Olomucine | CDK | 100 | ND | 0 |
| Roscovitine | CDK | 20 | 20 ± 5 | 0 |
| 5‐Iodotubercidin | ERK2, Adenosine Kinase, CKI, CK2II | 1 | 0 | ND |
| LFM‐A13 | BTK | 10 | 0 | 0 |
| SB‐202190 | P38 MAPK | 2 | ND | 0 |
| ZM336372 | cRAF | 5 | 0 | 0 |
| SU413 | Flk | 1 | 0 | 0 |
| GW5074 | cRaf | 1 | 0 | 0 |
| Palmytoil‐DL‐carmitine | PKC | 10 | ND | 70 ± 10 |
| Rottlerin | PKC delta | 10 | ND | 0 |
| Genistein |
Tyrosine Kinase |
1 | ND | 0 |
| Quercetin‐2H2O | PI 3‐K | 5 | 0 | 0 |
| Bay 11‐7082 | IKK pathway | 10 | ND | 0 |
| 5.6‐Dichloro‐1beta‐D‐ ribofuranosylbenzimidazole | CKII | 10 | 0 | 0 |
| 2,2,3,3’,4,4’‐Hexahydroxy‐1,1’‐biphenyl,‐6,6’.dimethanol dimethyl ether | PKC alpha, PKC delta | 1 | ND | 0 |
| SP 600125 | JNK | 5 | 0 | 0 |
| Indirubidin | GSK‐3 beta, CDK5 | 10 | ND | 0 |
| Indirubidin‐3’‐monoximine | GSK‐3 beta | 10 | 0 | ND |
| Y‐27632‐2HCl | ROCK | 0 | 0 | |
| Kenpaullone | GSK‐3 beta | 10 | 0 | 0 |
| Terreic acid | BTK | 0 | 0 | |
| Triciribine | Akt signaling pathway | 10 | 0 | 0 |
| BML‐257 | Akt | 10 | ND | 0 |
| SC‐514 | Ikk2 | 1 | ND | 0 |
| BML‐259 | CDK5/p25 | 0.5 | 0 | 0 |
| Apigenin | CK‐II | 20 | 0 | 0 |
| Rapamycin | mTor | 1 | 0 | 0 |
| AIP | CamK II | 1 | ND |
30 ± 10 70 ± 10 a |
The AIP inhibitory activity was also estimated by adding the compound to IVTT assay of S10A/G9R‐p27.