origene lentiviruses shctrl  (OriGene)

Journal: Cancers

Article Title: The RAL Small G Proteins Are Clinically Relevant Targets in Triple Negative Breast Cancer

doi: 10.3390/cancers16173043

Figure Lengend Snippet: Depletion of RALA or RALB reduces MDA-MB-468 tumor growth and is associated with changes in the tumor microenvironment. ( A ) Western blots displaying RALA and RALB expression in the MDA-MB-468 shRNA control (shCTRL), shRALA, and shRALB cells. ImageJ was used for quantification. ( B , C ) Effects of stable knockdown of RALA or RALB in MDA-MB-468 cells on 2D growth as measured by MTT (( B ), n = 4), and 3D growth as measured by GILA (( C ), n = 4) *, p < 0.05. ( D ) Stable knockdown of RALA or RALB had no effect on MDA-MB-468 cell migration (left: representative images, right: composite mean values, n = 2). ( E ) Comparison of tumor growth following orthotopic mammary fat pad injection of MDA-MB-468 shCtrl ( n = 20), shRALA ( n = 20), or shRALB ( n = 20). Results were combined from three independent experiments. ( F – H ) MDA-MB-468 shCTRL ( n = 9), shRALA ( n = 10), and shRALB ( n = 10) tumors IHC stained for ( F ) Ki-67, ( G ) CC3, or ( H ) CD31. ROIs were determined on each image separating the tumor from stroma before color deconvolution to extract DAB staining. A signal threshold (equivalent for each image) was then applied to the samples before measurement of the ROIs was performed to measure the % area of target staining in each region. Three representative photos from each sample were separately analyzed, and the mean values were used for comparisons among groups. *, p < 0.05. ( I ) MDA-MB-468 tumors stained with Masson’s Trichrome, denoted by white arrows, to analyze the collagen deposition in shCTRL ( n = 7), shRALA ( n = 8), or shRALB ( n = 9) MDA-MB-468 tumors. *, p < 0.05. ( J , K ) Graphs summarize the relative amounts of secreted proteins detected in the conditioned media from the MDA-MB-468 shCTRL, shRALA, and shRALB cultures.

Article Snippet: MDA-MB-468 parental cells were transduced with the Origene lentiviruses shCTRL (TR30021V), shRALA (TL309957VC, 5′-CTGGTTGGTAACAAATCAGATTTAGAAGA-3′), and shRALB (TL309956VD, 5′-GAACAGATTCTCCGTGTGAAGGCTGAAGA-3′).

Techniques: Western Blot, Expressing, shRNA, Control, Knockdown, Migration, Comparison, Injection, Staining

Journal: Cancers

Article Title: The RAL Small G Proteins Are Clinically Relevant Targets in Triple Negative Breast Cancer

doi: 10.3390/cancers16173043

Figure Lengend Snippet: Loss of RALA or RALB does not negatively impact SKBR3 tumor growth or viability. ( A ) Western blots displaying RALA and RALB expression in the SKBR3 shRNA control (shCTRL), shRALA, and shRALB cells. ImageJ was used for quantification. ( B ) Comparison of tumor growth following orthotopic mammary fat pad injection of SKBR3 shCTRL ( n = 10), shRALA ( n = 9), or shRALB ( n = 10). Results were from a single experiment. ( C – E ) Effects of stable knockdown of RALA or RALB in SKBR3 cells on 2D growth as measured by MTT (( C ), n = 5), 3D growth as measured by GILA (( D ), n = 5), and cell migration (( E ), n = 2). *, p < 0.05.

Article Snippet: MDA-MB-468 parental cells were transduced with the Origene lentiviruses shCTRL (TR30021V), shRALA (TL309957VC, 5′-CTGGTTGGTAACAAATCAGATTTAGAAGA-3′), and shRALB (TL309956VD, 5′-GAACAGATTCTCCGTGTGAAGGCTGAAGA-3′).

Techniques: Western Blot, Expressing, shRNA, Control, Comparison, Injection, Knockdown, Migration