Nc sirna sc 37007

Plasmids pCMV6-ddk-myc and pCMV6-ddk-myc-Lasp1 were purchased from Origene (RC219975, Rockville, MD, USA). Lasp1-siRNA (sc-105607) and NC-siRNA (sc-37007) were purchased from Santa Cruz Biotechnology. Transfection was carried out using the Lipofectamine 3000 reagent (Invitrogen) according to the manufacturer’s instructions.

Plasmids pCMV and pCMV‐Myc‐SP1 were purchased from Sino Biological (HG12024‐NM). Plasmid pcDNA3.1, pcDNA3.1‐BHLHE40 and pcDNA3.1‐BHLHE41 were generous gifts from Prof. Kijima, and the efficiency was demonstrated in a previous study.16 The plasmid pcDNA3.1‐BHLHE40 was inserted with N‐His‐Flag‐tag, and the plasmid pcDNA3.1‐BHLHE41 was inserted with N‐His‐Myc‐tag as described previously.25 The MCF‐7 cells were seeded at a density of 1 × 10⁵ cells per well (35 mm). After 24 hours, the cells were transfected with BHLHE40 or BHLHE41 using Lipofectamine LTX (Invitrogen). The cells were incubated for 24 hours following transfection and subjected to various analyses. NC‐siRNA (sc‐37007) was purchased from Santa Cruz Biotechnology. The sequences for the sense and antisense of three kinds of BHLHE41 siRNA are shown in Table 2. For the siRNA transfection experiments, MCF‐7 and MDA‐MB‐231 cells were seeded at 5 × 10⁴ cells per 35‐mm well. After 24 hours, the siRNA was transfected into the cells using Lipofectamine 3000 reagent (Invitrogen) according to the manufacturer's instructions. The transfected cells were incubated for another 48 hours and subjected to various analyses. The total amount of siRNA was adjusted with the control siRNA.

We bought the pCMV6-ddk-myc-SETD5 and pCMV6-ddk-myc plasmids from Origene (RC240118, Rockville, MD, USA). SETD5-siRNA (sc-78478) and NC-siRNA (sc-37007) were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Transfection was carried out using the Lipofectamine 3000 reagent (Invitrogen, Carlsbad, CA, USA) according to the manufacturer’s instructions.

BMDMs were seeded in 6-well-plates (2×10⁵ cells per well). After 24 h, they were transfected with either small interfering RNA (siRNA) against USP7 or negative control (NC) siRNA using Lipofectamine RNAiMAX (Invitrogen, Carlsbad, CA, USA). Cells were harvested after 48 h and processed for western blotting and flow cytometry. USP7 siRNA (sc-77373) and NC siRNA (sc-37007) were purchased from Santa Cruz Biotechnology, Inc.