Supplementary Figure 6 from Inactivation or Loss of TTP Promotes Invasion in Head and Neck Cancer via Transcript Stabilization and Secretion of MMP9, MMP2, and IL-6
Year: 2023
Type: supplementary-materials
Abstract: PDF file - 1486K, Supplementary Figure S6. Rap1B induces p38-mediated phosphorylation of TTP, which promotes secretion of pro-inflammatory mediators. (A) Whole cell lysates from UM-SCC-81B cells treated with IL-1β for 0, 2, 5, or 10 minutes were immunoblotted with phospho-p38(pp38), p38, rap1B and actin (loading control) antibodies. Active rap1B (rap1BGTP) was retrieved by a ralGDS pull-down assay. p38 was used for normalization of pp38 and rap1B was used for normalization of rap1BGTP. Signal intensity was quantified, normalized to its corresponding loading control and expressed as percent of time zero. Data are representative of three independent experiments. (B) UM-SCC-81B cells were transfected with two siRNAs targeting rap1B (si6 and si7) or NT and were stimulated with IL-1β or PBS (control) for 10 minutes. Whole cell lysates were immunoblotted with pp38, p38 and rap1B antibodies. Signal intensity was quantified, normalized to p38 and expressed as percent of stimulated NT. (C) UM-SCC-81B cells were transfected with sip38 or NT treated with PBS (control) or IL-1β for 10 minutes. Whole cell lysates were immunoprecipitated with TTP antibody bound AminoLink Plus Coupling Resin beads and eluted samples were immunoblotted with phosphoserine and TTP antibody. Signal intensity was quantified, normalized to input and expressed as percent of unstimulated NT. Whole cell lysates from NT and sip38 were also immunoblotted with pp38, p38 and actin antibodies. Signal intensity was quantified, normalized to p38 then actin and expressed as percent of unstimulated NT. Three independent experiments were performed. (D) UM-SCC-81B cells were transfected with sip38 or NT and whole cell lysates were immunoblotted for p38 and actin. Signal intensity was quantified, normalized to Actin and expressed as percent of NT. Interleukin-6 (IL-6) secretion was determined in conditioned media via ELISA with two replicates in each of three independent experiments. (*p<0.01).
Language: English
Subfield: Cancer Research
Domain: Life Sciences
Open Access status: gold
OpenAlex ID: W4361882807