control lentiviral particles shctrl  (Genechem)

Journal: Journal of Translational Medicine

Article Title: CCDC137 knockdown suppresses bladder cancer progression by downregulating SCD

doi: 10.1186/s12967-025-07033-w

Figure Lengend Snippet: Exploration of potential downstream mechanism and upstream regulators of CCDC137. A Heatmap shows DEGs identified by RNA sequencing in T24-shCtrl and T24-shCCDC137-1 cells. B Bar plot displays GO and KEGG functional enrichment results of downregulated DEGs from RNA sequencing. C GSVA-Hallmark pathway enrichment analysis displayed differences between CCDC137 positive (CCDC137 +) and CCDC137 negative (CCDC137 −) cells based on single-cell sequencing data. D mRNA and protein expression of stearoyl-CoA desaturase (SCD) were detected by qRT-PCR and Western blot. E DecoupleR was used to analyze differences in transcription factor activity between CCDC137 + and CCDC137 − epithelial cells in single-cell sequencing data. F TF-Target Finder was employed to identify potential upstream transcription factors of CCDC137. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001; ns, no statistical significance

Article Snippet: The shCCDC137 lentiviral particles and corresponding control lentiviral particles (shCtrl) were purchased from Genechem Co., Ltd (Shanghai, China).

Techniques: RNA Sequencing, Functional Assay, Sequencing, Expressing, Quantitative RT-PCR, Western Blot, Activity Assay

Journal: Journal of Translational Medicine

Article Title: CCDC137 knockdown suppresses bladder cancer progression by downregulating SCD

doi: 10.1186/s12967-025-07033-w

Figure Lengend Snippet: In vivo validation of CCDC137 regulating tumor growth. A Subcutaneous xenograft models were established in nude mice to validate the regulatory role of CCDC137 in tumor growth. B Tumor volume growth curves were plotted over 28 days after tumor cell inoculation. C Tumor weights of T24-shCtrl and T24-shCCDC137-1 groups were measured on day 28. D The Graphical Abstract described the key results in this study. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001; ns, no statistical significance

Article Snippet: The shCCDC137 lentiviral particles and corresponding control lentiviral particles (shCtrl) were purchased from Genechem Co., Ltd (Shanghai, China).

Techniques: In Vivo, Biomarker Discovery

Journal: BMC Cancer

Article Title: NSDHL contributes to breast cancer stem-like cell maintenance and tumor-initiating capacity through TGF-β/Smad signaling pathway in MCF-7 tumor spheroid

doi: 10.1186/s12885-024-13143-3

Figure Lengend Snippet: NSDHL knockdown suppresses tumor initiation and growth in tumor spheroid-injected xenograft tumor models. (A) Representative flow cytometry histogram of mCherry-positive cells sorted from NSDHL shRNA (shNSDHL) or control shRNA (shCtrl)-tagged mCherry-transduced MCF-7 cells. (B) Representative confocal images of mCherry-positive cells in shNSDHL- or shCtrl-spheroids. mCherry (red) and DAPI (blue). Scale bar: 100 μm C-E. NSDHL mRNA and protein levels and representative western blot images of NSDHL and mCherry in shNSDHL-MCF-7 spheroids relative to shCtrl-MCF-7 spheroids by qRT-PCR ( n = 6) and western blot analysis ( n = 6). F-G. Representative flow cytometry dot plots and quantification of CD44+/CD24 − cells in shCtrl and shNSDHL MCF-7 spheroids ( n = 3). H-I. Representative flow cytometry dot plots and quantification of ALDH + cells in shCtrl- and shNSDHL-MCF-7 spheroids using ALDEFLUOR assay ( n = 3). J. Tumor volume was measured weekly in shCtrl- or shNSDHL-MCF-7 spheroid-injected mice ( n = 6) for 56 days post-injection. K-L. Gross images and wet weights of tumors removed from shNSDHL- or shCtrl-MCF-7 spheroid-injected mice ( n = 6) at 56 days post-injection. M-N. Representative western blot images and quantification data of NSDHL proteins in shNSDHL-MCF-7 tumors compared to shCtrl-MCF-7 tumors ( n = 6). O. Representative NSDHL and mCherry immunohistochemistry images and H&E images of shNSDHL- or shCtrl-MCF-7 tumor tissues. Scale bar: 100 μm. The bar graph shows the mean ± standard deviation. * P < 0.05, ** P < 0.01, *** P < 0.001 as compared to shCtrl using unpaired t-tests

Article Snippet: shRNA lentiviral particles (shNSDHL) (LPP-HSH103352-LVRU6MH-c-100) and control shRNA lentiviral particles (shCtrl) (LPP-CSHCTR001-LVRU6MH-025-C) were purchased from GeneCopoeia (Rockville, MD, USA).

Techniques: Knockdown, Injection, Flow Cytometry, shRNA, Control, Western Blot, Quantitative RT-PCR, Immunohistochemistry, Standard Deviation

Journal: BMC Cancer

Article Title: NSDHL contributes to breast cancer stem-like cell maintenance and tumor-initiating capacity through TGF-β/Smad signaling pathway in MCF-7 tumor spheroid

doi: 10.1186/s12885-024-13143-3

Figure Lengend Snippet: Incidence of palpable tumors in mice injected with shCtrl or shNSDHL cells

Article Snippet: shRNA lentiviral particles (shNSDHL) (LPP-HSH103352-LVRU6MH-c-100) and control shRNA lentiviral particles (shCtrl) (LPP-CSHCTR001-LVRU6MH-025-C) were purchased from GeneCopoeia (Rockville, MD, USA).

Techniques: Injection

Journal: BMC Cancer

Article Title: NSDHL contributes to breast cancer stem-like cell maintenance and tumor-initiating capacity through TGF-β/Smad signaling pathway in MCF-7 tumor spheroid

doi: 10.1186/s12885-024-13143-3

Figure Lengend Snippet: NSDHL knockdown reduced the BCSC population with CD44+/CD24 − phenotype and ALDH activity in tumor spheroid-injected xenograft tumor tissues, accompanied by decreased Smad2/3 phosphorylation and SOX2 expression. A-D. Representative flow cytometry histograms, dot plots, and quantification of CD44 + and CD24- cells in control (shCtrl) and NSDHL shRNA (shNSDHL)-transduced MCF-7 tumors ( n = 4). E-F. Representative flow cytometry dot plots and quantification of ALDH + cells in shCtrl- and shNSDHL MCF-7 tumors using ALDEFLUOR assay ( n = 4). G. Representative double immunohistochemistry images of CD44 + and ALDH1A1 + cells in shCtrl and shNSDHL MCF-7 tumors. CD44 (green), ALDH1A1(red). Double CD44+/ALDH1A1 + cells (arrow). H-O. Representative western blot images and quantification data of phopho-Smad2 and 3, Smad2, Smad3, SOX2, and NANOG levels in shNSDHL MCF-7 tumors relative to shCtrl MCF-7 tumors ( n = 5). All graphs show the mean ± standard deviation. * P < 0.05, compared with shCtrl using unpaired t-tests

Article Snippet: shRNA lentiviral particles (shNSDHL) (LPP-HSH103352-LVRU6MH-c-100) and control shRNA lentiviral particles (shCtrl) (LPP-CSHCTR001-LVRU6MH-025-C) were purchased from GeneCopoeia (Rockville, MD, USA).

Techniques: Knockdown, Activity Assay, Injection, Expressing, Flow Cytometry, Control, shRNA, Immunohistochemistry, Western Blot, Standard Deviation

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

Journal: BMC cancer

Article Title: NAD(P)-dependent steroid dehydrogenase-like is involved in breast cancer cell growth and metastasis.

doi: 10.1186/s12885-020-06840-2

Figure Lengend Snippet: Fig. 2 NSDHL knockdown decreases viability, proliferation, and colony and sphere formation abilities of BT-20 and MDA-MB-231 cells. a, b Data of cell viability and cell cycle in BT-20 and MDA-MB-231 cells transfected with NSDHL siRNA or control siRNA (20 nM); c, d Representative images and data analyzed in colony formation of BT-20 and MDA-MB-231 cells transfected with NSDHL siRNA or control siRNA (20 nM); e Representative image and data analyzed in 3D sphere formation of BT-20 and MDA-MB-231 cells transfected with NSDHL siRNA or control siRNA (20 nM). All data represent the means ± standard deviations of three independent experiments, each performed in triplicates. *p < 0.05, **p < 0.01, ***p < 0.001

Article Snippet: Short hairpin RNA (shRNA) lentiviral transduction Precisely, 4 × 105 MDA-MB-231 cells were seeded in each well of 12-well plate 24 h prior to viral infection and were replaced with media containing 5 μg/ml polybrene® (sc-134220, Santa Cruz) and 20 μl of NSDHLtargeting shRNA lentiviral particles (shNSDHL) (sc90849-V, Santa Cruz) or control shRNA lentiviral particles (shCtrl) (sc-108080, Santa Cruz).

Techniques: Knockdown, Transfection, Control

Journal: BMC cancer

Article Title: NAD(P)-dependent steroid dehydrogenase-like is involved in breast cancer cell growth and metastasis.

doi: 10.1186/s12885-020-06840-2

Figure Lengend Snippet: Fig. 3 NSDHL knockdown decreases the migration and invasion abilities of BT-20 and MDA-MB-231 cells. a,b,c Representative images and data analyzed in transwell migration assay, invasion assay and, wound healing assay of BT-20 and MDA-MB-231 cells transfected with NSDHL siRNA or control siRNA (20 nM). Data represent the means ± standard deviations of three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001

Article Snippet: Short hairpin RNA (shRNA) lentiviral transduction Precisely, 4 × 105 MDA-MB-231 cells were seeded in each well of 12-well plate 24 h prior to viral infection and were replaced with media containing 5 μg/ml polybrene® (sc-134220, Santa Cruz) and 20 μl of NSDHLtargeting shRNA lentiviral particles (shNSDHL) (sc90849-V, Santa Cruz) or control shRNA lentiviral particles (shCtrl) (sc-108080, Santa Cruz).

Techniques: Knockdown, Migration, Transwell Migration Assay, Invasion Assay, Wound Healing Assay, Transfection, Control

Journal: BMC cancer

Article Title: NAD(P)-dependent steroid dehydrogenase-like is involved in breast cancer cell growth and metastasis.

doi: 10.1186/s12885-020-06840-2

Figure Lengend Snippet: Fig. 4 NSDHL knockdown suppresses total cholesterol level and promotes erlotinib response in MDA-MB-231 cell. a Total cholesterol levels measured in BT-20 and MDA-MB-231 cells transfected with NSDHL siRNA or control siRNA (20 nM); b Dose-response curve of erlotinib in BT-20 and MDA-MB-231 cells transfected with NSDHL siRNA or control siRNA (20 nM); c Representative western blot images of NSDHL, EGFR, and precursor and mature SREBP-1 and data of relative expression levels of NSDHL, EGFR, and precursor SREBP-1 in BT-20 and MDA-MB-231 cells transfected with NSDHL siRNA or control siRNA (20 nM). Data represent the mean ± standard deviation of three independent experiments. *p < 0.05, ***p < 0.001

Article Snippet: Short hairpin RNA (shRNA) lentiviral transduction Precisely, 4 × 105 MDA-MB-231 cells were seeded in each well of 12-well plate 24 h prior to viral infection and were replaced with media containing 5 μg/ml polybrene® (sc-134220, Santa Cruz) and 20 μl of NSDHLtargeting shRNA lentiviral particles (shNSDHL) (sc90849-V, Santa Cruz) or control shRNA lentiviral particles (shCtrl) (sc-108080, Santa Cruz).

Techniques: Knockdown, Transfection, Control, Western Blot, Expressing, Standard Deviation

Journal: BMC cancer

Article Title: NAD(P)-dependent steroid dehydrogenase-like is involved in breast cancer cell growth and metastasis.

doi: 10.1186/s12885-020-06840-2

Figure Lengend Snippet: Fig. 5 NSDHL knockdown suppressed tumor growth and lung metastasis of MDA-MB-231 xenograft mice. a, b Data of relative NSDHL mRNA level analyzed by real-time RT-PCR and representative western blot images of NSDHL in MDA-MB-231 cells transduced with NSDHL shRNA or control shRNA lentivirus; c Data of tumor volume measured weekly in NSDHL shRNA or control shRNA mice; d Gross images and wet weight of tumors removed from NSDHL shRNA or control shRNA mice at 44 days post-injection; e Representative NSDHL immunohistochemistry image and scores analyzed from NSDHL shRNA or control shRNA tumor tissues; f Gross and H&E images of lungs and data of metastatic foci analyzed from NSDHL shRNA or control shRNA lung tissues. In vitro data represent the means ± standard deviations of three independent experiments. Animal data represent the means ± standard deviations of five mice per group. *p < 0.05, **p < 0.01

Article Snippet: Short hairpin RNA (shRNA) lentiviral transduction Precisely, 4 × 105 MDA-MB-231 cells were seeded in each well of 12-well plate 24 h prior to viral infection and were replaced with media containing 5 μg/ml polybrene® (sc-134220, Santa Cruz) and 20 μl of NSDHLtargeting shRNA lentiviral particles (shNSDHL) (sc90849-V, Santa Cruz) or control shRNA lentiviral particles (shCtrl) (sc-108080, Santa Cruz).

Techniques: Knockdown, Quantitative RT-PCR, Western Blot, Transduction, shRNA, Control, Injection, Immunohistochemistry, In Vitro