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Additional file 1 of Revealing RCOR2 as a regulatory component of nuclear speckles

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posted on 25.11.2021, 05:14 by Carlos Rivera, Daniel Verbel-Vergara, Duxan Arancibia, Anna Lappala, Marcela González, Fabián Guzmán, Gianluca Merello, Jeannie T. Lee, María Estela Andrés
Additional file 1: Figure S1. Anti-RCOR2 antibody validation. (A) Western blot of total extracts from N2A, HEK293T and PC12 cells using the anti-RCOR2 antibody. (B) HEK293T cells were transfected to ectopically overexpress RCOR2-Myc. Western blot was performed using the anti-RCOR2 antibody. β-Actin was assayed as a loading control. (C) Confocal image showing immunostaining of anti Myc-epitope (red) and anti-RCOR2 (green) on transiently transfected PC12 cells with RCOR2-Myc. (D) Peptide competition assay. RCOR2 immunostaining was performed on HT22 cells. Anti-RCOR2 antibody was pre-incubated with RCOR2 (2–61) peptide or H3 (1–20) peptide (negative control) at 1:1 and 1:5 antibody:peptide molar ratios, respectively. (E) Western blot analysis of RCOR2 after HEK293T cells were transfected with siRNA targeting RCOR2. β-Tubulin was assayed as loading control. (F) RCOR2 immunofluorescence was performed on HT22 cells that were transduced with lentiviral particles to perform shRNA mediated knockdown of RCOR2. Fluorescence intensity was quantitated on the right plot.


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