Journal: Cancer research

Article Title: Targeting the HuR oncogenic role with a new class of cytoplasmic dimerization inhibitors

doi: 10.1158/0008-5472.CAN-20-2858

Figure Lengend Snippet: (A) Graphs illustrate the percent reduction of Bcl2/18S and Mcl1/18S mRNAs following cell treatment with lead compounds versus control (vehicle treatment). Results are shown as mean±SD (n=3), differences statistically significant for all cell lines with P<0.005, Student t-test. (B) Representative western blots confirm the reduction of Bcl2 and Mcl1 proteins in cytoplasmic fractions of U251 and XD456 cell lines following treatment with lead compounds, 10 uM for 48 hours. SOX2 reduction in nuclear fractions indicates a decrease in tumorigenicity of cell lines after treatment with lead compounds. (C) Representative western blots illustrate appearances of cleaved PARP and cleaved caspase-3 in glioma cell lines after treatment with lead compounds, 10 uM for 48 hours. (D) Graphs illustrate a significant reduction of colony formations in U251 and XD456 cell lines after SRI-42127 (3 uM and 10 uM treatments) versus control (vehicle treatment). Results are shown as mean±SD, (n=3) and (n=4) for attached and anchorage-independent assays, respectively; P<0.005, Student t-test for all assays. Examples of colony formations are shown in the U251 cell line.

Article Snippet: The quantifications of Bcl2, Mcl1, and 18S transcripts were performed by using Taqman technique with the following gene-specific probes: Hs00153350_m1, Hs03003631_g1, and Hs00766187_m1 (ThermoFisher Scientific) for Bcl2, 18S, and Mcl1, respectively ( 19 ). mRNA isolation from adherent cell culture mRNA isolation and purification from adherent cells were performed as previously described by using the RNeasy Mini Kit and QIAshredder columns (Qiagen, Valencia, CA) ( 19 ).

Techniques: Control, Western Blot

Journal: Cancer research

Article Title: Targeting the HuR oncogenic role with a new class of cytoplasmic dimerization inhibitors

doi: 10.1158/0008-5472.CAN-20-2858

Figure Lengend Snippet: (A-D) SRI-42127 suppresses glioma progression in vivo. Mice were randomly divided into two groups after four and half days of the intracranial tumor establishment and treated with vehicle (control group) or SRI-42127 compound (SRI-42127 group), 15 mg/kg, twice a day for three weeks via intraperitoneal injection. Representative scans illustrate the tumor reporter images superimposed with mouse images; luminescence/color scales are beside the corresponding scans. Graphs illustrate luminescence signals from intracranial tumors on the second (top) and third (bottom) weeks of treatment; results are shown as mean±SD (radiance, p/sec/cm2/sr); the differences between groups were not statistically significant, P=0.07 (n=5) for the second week, P=0.069 (n=5) for the third week, Student t-test. The weight changes in group treated with SRI-42127 compound were not significant compared to the control group (P>0.05, Student t-test). Immunostaining for HuR (B), Bcl2 (C), and MCl1 (D) on tumor brain tissue from control and treated with SRI-42127 compound mouse groups. (E). Computational docking of SRI-42127 at HuR. SRI-42127 compound and key residues of HuR are represented with green and purple carbons, respectively. Hydrogen bonds, hydrophobic contacts, and pi-pi stacking (or pi-cation interactions) are indicated by cyan, orange, and green dash lines, respectively. Grey and blue ribbons represent RRM1 and RRM2 domains, respectively.

Article Snippet: The quantifications of Bcl2, Mcl1, and 18S transcripts were performed by using Taqman technique with the following gene-specific probes: Hs00153350_m1, Hs03003631_g1, and Hs00766187_m1 (ThermoFisher Scientific) for Bcl2, 18S, and Mcl1, respectively ( 19 ). mRNA isolation from adherent cell culture mRNA isolation and purification from adherent cells were performed as previously described by using the RNeasy Mini Kit and QIAshredder columns (Qiagen, Valencia, CA) ( 19 ).

Techniques: In Vivo, Control, Injection, Immunostaining