Ab133354

HEK293T/17 were prepared as described above for RNA FISH. After the cells were permeabilized, the primary antibodies, anti-hnRNP K mouse monoclonal antibody [3C2]-ChIP Grade (ab39975, Abcam) and anti-PTBP1 mouse monoclonal antibody (32-4800, Thermo Fisher Scientific), were added and hybridized overnight. Alexa Fluor 647–conjugated goat anti-mouse IgG secondary antibodies (A-21236, Thermo Fisher Scientific) were used to visualize the results. Several organelle marker antibodies were tested as follows; anti-SC35 (S4045, Sigma-Aldrich), anti-nmt55/p54nrb (NONO, ab70335, Abcam), anti-ILF3 (ab133354, Abcam), anti-EF1A (sc-21758, SantaCruz), anti-ATP5 (mitochondria, ab14748, Abcam), anti-Calnexin (endoplasmic reticulum, ab202572, Abcam) and J2 monoclonal antibody (dsRNA, 10010200, SCICONS). MALAT1 probes labeled with Atto 633 (Supplementary Table S2).

Cultured cells and tissues were lysed with RIPA buffer (Beibo, China) and then protein for further western blot following the protocol as describe previously. The antibodies were used in present study as following: anti-ERp57 (Abcam, ab13506), anti-ILF3 (Abcam, ab133354), anti-Cyclin E1 (Abcam, ab33911), anti-Cyclin D1 (Proteintech, 26,939–1-AP), anti-STAT3 (Abcam, ab68153), anti-pSTAT3 (Abcam, ab76315) and anti-β-actin (Abcam, ab6276).

RIPA buffer (Beyotime Biotechnology, Shanghai, China) was used to lysis cells and isolated proteins. Extracted proteins were then quantified with a BCA Protein assay kit (Beyotime Biotechnology) according to the product specification. The proteins were separated with electrophoresis, then transferred from the gel to a polyvinylidene difluoride (PVDF) membrane using the semi-dry-transfer method. After transfer, the membrane was washed with TBST buffer twice and blocked at 25 °C with 5% defatted milk powder for 1 h, and the membrane was washed with TBST three times and put into a sealed bag containing the primary antibody solution (1:1000) and incubated overnight at 4 °C. Next day, the membranes were hybridized with specific HRP-conjugated secondary antibodies at ~25°C for 1 h with a dilution ratio of 1:5000. Imagines were captured by using ChemiDoc^TM MP Imaging System (Bio-Rad). Primary antibodies used in this study include Vimentin (5741 S, CST), Snail (3879 S, CST), ILF3 (ab133354, Abcam), METTL3 (96391, CST), FTO (31687 S, CST), HA-tag (3724 S, CST), TGF beta 2 (36495, Abcam), N-cadherin (13116 S, CST), CD9 (13403, CST), CD63 (ab134045, Abcam), E-cadherin (3195 S, CST), TSG101 (ab125011, Abcam) and anti-GAPDH (5174 S, CST). Non-specific bands may exist to mix the visualization of ILF3, and all experimental operations will be performed accurately to quantify correctly.