LTβR-IN-1 is a potent, selective lymphotoxin β receptor (LTβR) inhibitor. LTβR-IN-1 also selectively inhibits the nuclear translocation of p52 depended on TNF12A, instead of the nuclear translocation of p65 mediated by TNF-α receptor. LTβR-IN-1 regulates the NF-kB signaling pathway IN a ligand-independent manner In Vitro LTβR-IN-1 (compound 919278) (1 nM-100 μM; 30 min) inhibits p52 nuclear translocation in response to stimulation with Anti-LTβR or TWEAK (20 ng/mL, respectively; 4 h) with IC 50 s of 0.169 μM and 0.167 μM, respectively. LTβR-IN-1 (3 nM-30 μM; 45 min) reduces the binding affinities of both CDK12 and its associated protein, CCNK in both TWEAK-stimulated and control U-2 OS cells, with IC 50 s of 50-61 nM for CDK12 and 29-68 nM for CCNK, respectively. LTβR-IN-1 reduces the mRNA abundance of MAP3K14 in with an IC 50 value of 0.32 μM in U-2 OS cells. LTβR-IN-1 (1-10 μM; 30 min) decreases the phosphorylation of serine-2 (Ser2) on the C-terminal domain (CTD) of RNA polymerase (Pol) II. LTβR-IN-1 regulates the noncanonical pathway in a ligandindependent manner and selectively inhibits the noncanonical pathway while sparing the canonical NF-kB signaling pathway. MCE has not independently confirmed the accuracy of these methods. They are for reference only. Western Blot AnalysisCell Line: U-2 OS cells Concentration: 1 μM, 3 μM, and 10 μM Incubation Time: 30 min Result: Reduced the phosphorylating of Ser2 in cells stimulated by 20 ng/mL TWEAK for 4 hr. Note: CDK12 activated RNA Pol II–mediated transcription by phosphorylating Ser 2 within the 52 heptad (Y1S2P3T4S5P6S7) repeats in the C-terminal domain (CTD) of RNA Pol II. Ser 2 phosphorylation aids in the release of paused RNA Pol II from promoters, resulting in transcriptional elongation, which is particularly important for the transcription of some long, complex genes. Form:Solid IC50& Target:LTβR,p52 translocation,MAP3K14,NF-κB