Bovine serum albumin fraction 5

Bovine Serum Albumin-Fraction V was purchased from HiMedia Laboratories GmbH, Germany). Sodium phosphate, sodium chloride, and sodium acetate were purchased from Sigma Aldrich (St. Louis, USA). Scramble (Silencer) 5′-FAM labelled siRNA (sense: 5′-UUCUCCGAACGUGUCACGUdTdT-3′; antisense: 5′-ACGUGACACGUUCGGAGAAdTdT-3′), HDAC4 siRNA (sense: 5′-GGUGCUUAUGGAAAGGGAUTT-3′; antisense: 5′-AUCCCUUUCCAUAAGCACCTT-3′), bicinchoninic acid (BCA) protein assay kit and MTT reagent (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide) was acquired from Thermo Fisher Scientific (Massachusetts, USA). DEPC treated RNase free water was used for further experimentation with siRNA and procured from Invitrogen (Thermo scientific, Massachusetts, USA). All other reagents and solvents were of analytical grade unless otherwise specified. Agarose and sodium chloride were obtained from Sigma Aldrich (Mumbai, India). RPMI 1640 media, fetal bovine serum (FBS), trypsin-EDTA, penicillin-streptomycin and 1x ITS (Insulin-Transferrin-Selenium) were obtained from Invitrogen and Gibco (Invitrogen, California, USA, Gibco, Life Technologies, Grand Island, USA). Opti-MEM media, Hoechst 33342 were obtained from Gibco, (Life Technologies, Grand Island, USA).

Human cervical cancer cell lines with known HPV positivity for HPV type 16 – SiHa, HPV type 18 – HeLa; and HPV-negative C33a were originally procured from ATCC. The materials used in the study have been listed along with their source of procurement. The materials used in the study have been listed along with their source of procurement. DMEM (#AL111-18X500ML), MEM (#AT154), antibiotic solutions (#A018), bovine serum albumin fraction V (#RM10409) were procured from HiMedia Laboratories Pvt. Ltd. (Mumbai, India), Pierce™ BCA Protein Assay Kit (#23225), exosome-depleted serum One Shot™ format (#A2720803), Precision Plus Protein Dual Color Standards from Bio-Rad, USA (#161-0374), High-Capacity cDNA Reverse Transcription Kit (Applied Biosystems™; #4368814), TRIzol RNA Isolation Reagent (#AM9738) were procured from Thermo Fischer Scientific (Waltham, USA). ExoEnrich™ (#PEC-50), ExoLyseP™ (#PEL-25P) were purchased from ExoCan Healthcare Technologies Ltd. (Pune, India). ECL-substrate (#SC-2048) from Santa Cruz Biotechnology, Inc. (Dallas, USA). All the antibodies were procured from Santacruz Biotechnology, Inc. (USA) and Sigma (St. Louis, USA) (Supplementary Table 1) and oligos used in the study were procured from Eurofins scientific (Supplementary Table 2). Millipore PVDF membrane (#HVLP04700), RNAse A (#P4170), and all other reagents unless specified were procured from Sigma.

The cells were harvested after 48 h of transfection using the mammalian protein extraction reagent (mPER; Thermo Fisher Scientific) supplemented with protease inhibitor cocktail (PIC, Thermo Fisher Scientific). Bicinchoninic acid (BCA) assay was performed for the estimation of protein concentration. Total cellular protein (40 μg per well) was run in the SDS-PAGE gels, transferred onto a PVDF membrane, blocked using bovine serum albumin fraction-V (HiMedia) and were incubated overnight with BANF1 (Abcam, Cambridge, UK), HBx (Santa Cruz Biotechnology, Dallas, TX, USA), or βactin (Santa Cruz). PVDF membranes were washed 6 × 5 min each and incubated with HRP-conjugated corresponding secondary antibodies and developed using ECL kit (Thermo Fisher Scientific) on photosensitive films.