Fig. 2: MACC1-AS1 directly interacts with STK33 to induce gemcitabine resistance.

A LC-MS/MS analysis was used to evaluate the downstream target via MACC1-AS1 overexpression PANC-1 cell, accompanied by vector cells. At the same time, KEGG pathway and cell components were put into work to analyze the results and to select the target. B Cell lysates of STK33 or purified His-tagged STK33 was pulled down by biotin-labeled MACC1-AS1 but not by MACC1-AS1 sense RNA (S, sense. AS, antisense). C RIP assays of MACC1-AS1, precipitated with STK33 in whole-cell lysates, and the RNA levels of MACC1-AS1 and β-actin were measured by qPCR analysis. D MACC1-AS1-binding proteins were tested by Western blot analysis, using anti-FLAG antibody affinity agarose beads and specific antibodies to identify the targets. E, F The full length of MACC1-AS1 and the T1, T2, T3 fragments were selected for further research from the predictive website, and RNA pull-down and Western blot assays were performed using synthesized FL and specific fragment of MACC1-AS1, which were incubated with cell lysates or purified His-tagged recombinant STK33. G CLIP-qPCR research of T1/T2/T3 fragment of MACC1-AS1 for STK33 binding was conducted. H RNA pull-down assays was used to confirm the MACC1-AS1 T3 fragment, interacting with STK33. I–K The interaction map, IF and Alphafold database were conducted to show the interaction fraction, cell co-location and specific part (K355) of STK33, which binds with MACC1-AS1 (T3 fragment).