CD74 siRNA and shRNA Plasmids (h)

Protocol tips

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	HT-1376 (G3) cell lines were purchased from the Type Culture Collection of Chinese Academy of Sciences (Shanghai, China). HT-1376 were all cultured in RPMI-1640 (Hyclone; GE Healthcare Life Sciences) medium supplemented with 10% fetal bovine serum (Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA). All cells were routinely cultured in a 37°C humidified incubator with 5% CO2 and 95% O2. The CD74 shRNA lentiviral particles (cat. no., sc-35023-V; 1.0×106/200 µl; Santa Cruz Biotechnology, Inc.) targeting the human CD74 transcript and control shRNA lentiviral particles (cat. no., sc-108080; 1.0×106/200 µl; Santa Cruz Biotechnology, Inc.) were used to knock down CD74 in HT-1376 cells. Cells were seeded at 2×105 cells/well in 6-well plates (Corning Incorporated, Corning, NY, USA) and grown to 60% confluence on the day of transfection. The media was removed from the plate wells and replaced with 2 ml complete medium containing Polybrene (cat. no., sc-134220; Santa Cruz Biotechnology, Inc.) at a final concentration of 5 µg/ml. Cells were transfected with control (scramble, 20 µl) or CD74 shRNA lentiviral particles (20 µl) diluted in media according to the manufacturer's protocol in 48 h. Infected cells were selected with puromycin (5 µg/ml; cat. no., 11811-023; Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA). A total of 72 h after transfection, transfected cells were verified by RT-PCR and western blotting analysis, comparing untreated and scramble cells.

Protocol tips

HT-1376 (G3) cell lines were purchased from the Type Culture Collection of Chinese Academy of Sciences (Shanghai, China). HT-1376 were all cultured in RPMI-1640 (Hyclone; GE Healthcare Life Sciences) medium supplemented with 10% fetal bovine serum (Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA). All cells were routinely cultured in a 37°C humidified incubator with 5% CO2 and 95% O2. The CD74 shRNA lentiviral particles (cat. no., sc-35023-V; 1.0×106/200 µl; Santa Cruz Biotechnology, Inc.) targeting the human CD74 transcript and control shRNA lentiviral particles (cat. no., sc-108080; 1.0×106/200 µl; Santa Cruz Biotechnology, Inc.) were used to knock down CD74 in HT-1376 cells. Cells were seeded at 2×105 cells/well in 6-well plates (Corning Incorporated, Corning, NY, USA) and grown to 60% confluence on the day of transfection. The media was removed from the plate wells and replaced with 2 ml complete medium containing Polybrene (cat. no., sc-134220; Santa Cruz Biotechnology, Inc.) at a final concentration of 5 µg/ml. Cells were transfected with control (scramble, 20 µl) or CD74 shRNA lentiviral particles (20 µl) diluted in media according to the manufacturer's protocol in 48 h. Infected cells were selected with puromycin (5 µg/ml; cat. no., 11811-023; Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA). A total of 72 h after transfection, transfected cells were verified by RT-PCR and western blotting analysis, comparing untreated and scramble cells.

Publication protocol

HT-1376 (G3) cell lines were purchased from the Type Culture Collection of Chinese Academy of Sciences (Shanghai, China). HT-1376 were all cultured in RPMI-1640 (Hyclone; GE Healthcare Life Sciences) medium supplemented with 10% fetal bovine serum (Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA). All cells were routinely cultured in a 37°C humidified incubator with 5% CO2 and 95% O2. The CD74 shRNA lentiviral particles (cat. no., sc-35023-V; 1.0×106/200 µl; Santa Cruz Biotechnology, Inc.) targeting the human CD74 transcript and control shRNA lentiviral particles (cat. no., sc-108080; 1.0×106/200 µl; Santa Cruz Biotechnology, Inc.) were used to knock down CD74 in HT-1376 cells. Cells were seeded at 2×105 cells/well in 6-well plates (Corning Incorporated, Corning, NY, USA) and grown to 60% confluence on the day of transfection. The media was removed from the plate wells and replaced with 2 ml complete medium containing Polybrene (cat. no., sc-134220; Santa Cruz Biotechnology, Inc.) at a final concentration of 5 µg/ml. Cells were transfected with control (scramble, 20 µl) or CD74 shRNA lentiviral particles (20 µl) diluted in media according to the manufacturer's protocol in 48 h. Infected cells were selected with puromycin (5 µg/ml; cat. no., 11811-023; Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA). A total of 72 h after transfection, transfected cells were verified by RT-PCR and western blotting analysis, comparing untreated and scramble cells.

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