Journal: Nature Communications

Article Title: KK-LC-1 as a therapeutic target to eliminate ALDH + stem cells in triple negative breast cancer

doi: 10.1038/s41467-023-38097-1

Figure Lengend Snippet: a Volcano plot showing up-regulated genes in docetaxel-resistant MDA-MB-231 cells (DTX_R) compared to their parental cells (Parental) (Log2 Fold change > 6, adj P value < 0.05, P values were determined using moderated t -statistic, adj P -value was calculated using Benjamini and Hochberg’s method and tails were two-sided). b Up-regulated genes in docetaxel-resistant MDA-MB-231 cells were intersected with the up-regulated genes in TNBC , . Expression levels of two genes were significantly higher in docetaxel-resistant MDA-MB-231 cells. c Heatmap showing expression of the two highly expressed genes. d Schematic showing the identification and isolation of primary ALDH + and ALDH - cells. e Among the two highly expressed genes, KK-LC-1 had the highest expression in ALDH + cells compared to ALDH − cells and was selected as a potential candidate that may regulate ALDH + cells in TNBC; data are presented as mean ± SD from three biologically independent experiments, statistical significance was determined using Student’s t -test and was two-sided. f, g : KK-LC-1 knockout significantly reduced the proportion of ALDH + cells and ALDH + CD44 high CD24 low cells in MDA-MB-231 cells; data are presented as mean ± SD from three biologically independent experiments and statistical significance was determined using Student’s t -test and was two-sided. h The proportion of ALDH + cells was significantly lower in ALDH + sh- KK-LC-1 #1 cells compared to ALDH + NC cells after 2 weeks in culture; data are presented as mean ± SD from three biologically independent experiments, statistical significance was determined using Student’s t -test and was two-sided. i A significantly higher proportion of ALDH + cells was derived from CD44 high CD24 low KK-LC-1 +/+ MDA-MB-231 compared to CD44 high CD24 low KK-LC-1 −/− MDA-MB-231 cells after 2 weeks in culture; data are presented as mean ± SD from three biologically independent experiments, statistical significance was determined using Student’s t -test and was two-sided. Source data are provided as a Source data file.

Article Snippet: MDA-MB-231 cells and MDA-MB-468 cells were resuspended in PBS and stained with phycoerythrin (PE)-conjugated anti-CD24 at a dilution of 1:100 (Cat: 130-112-656, Miltenyi Biotec) and allophycocyanin (APC)-conjugated anti-CD44 at a dilution of 1:500 (Cat: 12211-MM02-A, SinoBiological) for 30 min at 4 °C.

Techniques: Expressing, Isolation, Knock-Out, Derivative Assay

Journal: Life Science Alliance

Article Title: Dendritic cells maintain anti-tumor immunity by positioning CD8 skin-resident memory T cells

doi: 10.26508/lsa.202101056

Figure Lengend Snippet: (A) Representative flow plots showing expression of CD103 and CD69 on CD8 T cells isolated from LN and skin from melanoma-associated vitiligo (MAV)–affected mice. Cells were isolated from experimental mice 35 post-surgery and were identified by CD8 + CD62L neg CD44 + . (B) Skin from MAV-affected mice, stains identify CD8β (red), CD103 (green), and KLRG1 (white). Scale bar, 50 μm. (C) Number of CD103 neg KLRG1 neg CD8 + , CD103 + KLRG1 neg CD8 + , and CD103 neg KLRG1 + CD8 + T cells per cluster in a 10 μm thick skin section. Symbols represent individual clusters; horizontal lines indicate mean; n = 9 clusters. (D) Heat map of skin-resident memory transcripts comparing bulk endogenous CD8 T cells to Ag-specific Pmel CD8 T cells and naïve CD8 T cells. Bulk endogenous and Ag-specific Pmel CD8 T cells were isolated from MAV-affected skin, naïve CD8 T cells were isolated from spleen and LN of naïve Pmel mice.

Article Snippet: Antibodies from BioLegend: anti-mouse CD45-APC/Fire 750 (clone 30-F11; #147713), anti-mouse CD8α-PerCP/Cynine5.5, -PE/Cy7 (clone 53-6.7; #100733, #100721), anti-mouse Thy1.1-PerCP/Cynine5.5 (clone OX-7; #202515), anti-mouse CD103-Alexa Fluor 647, -FITC (clone 2E7; #121409, #121419), anti-mouse CD44-APC/Fire 750 (clone IM7; #103061), anti-mouse CD62L-Brilliant Violet 510 (clone MEL-14; 104441), anti-mouse CXCR6-Brilliant Violet 421 (clone SA051D1; #151109), anti-mouse CD11c Alex Fluor 647 (clone N418; #117314), anti-mouse CD11b-Alexa Fluor 488 (clone M1/70; #101219); Bioss: anti-mouse CXCL16 Alexa Fluor 488 (polyclonal; #bs-1441R-A488).

Techniques: Expressing, Isolation

Journal: Life Science Alliance

Article Title: Dendritic cells maintain anti-tumor immunity by positioning CD8 skin-resident memory T cells

doi: 10.26508/lsa.202101056

Figure Lengend Snippet: (A) Experimental outline to induce MAV. Mice received 5 × 10 5 activated Pmel CD8 T cells expressing a luciferase reporter gene (Luc + Pmel) 5 d prior to tumor resection. (B) Luminescence signal of Luc + Pmel cells over the course of 21 d post tumor excision in unaffected and MAV-affected mice. (C) Representative flow plots showing expression of CXCR6 and CD44 on Luc + Pmel CD8 T cells. Cells were isolated from experimental mice on days 7, 14, or 35 post-surgery and were identified by CD8 + Thy1.1 + CD44 + ; Naïve Pmel CD8 T cells were isolated from naïve mice and identified by CD8 + Thy1.1 + CD44 − . Graphs are compiled data comparing the percentage of CXCR6 expression on CD8 + Thy1.1 + CD44 + (days 7, 14, and 35) or CD8 + Thy1.1 + CD44 − (naïve) cells isolated from skin, skin-draining LN, or spleen. (D) CXCR6 mean fluorescence intensity expression on CD8 + Thy1.1 + CD44 + (day 7, 14, 35) or CD8 + Thy1.1 + CD44 − (naïve) cells isolated from skin, skin-draining LN, or spleen. (E) Colocalization of CD8 and CXCR6 staining in skin from MAV-affected patients. Stains identify CD8 (green) and CXCR6 (pink); dark red indicates colocalization of CD8 and CXCR6. Scale bar, 25 μm. (F) Percent of CD8 T cells in skin from MAV-affected patients that are positive for CXCR6. (C, D, F) Symbols represent individual mice (C, D), individual patients (F); horizontal lines indicate mean. (C, D) n = 2–4; representative of three independent experiments (C, D). Significance was determined by one-way ANOVA; ** P ≤ 0.010.

Article Snippet: Antibodies from BioLegend: anti-mouse CD45-APC/Fire 750 (clone 30-F11; #147713), anti-mouse CD8α-PerCP/Cynine5.5, -PE/Cy7 (clone 53-6.7; #100733, #100721), anti-mouse Thy1.1-PerCP/Cynine5.5 (clone OX-7; #202515), anti-mouse CD103-Alexa Fluor 647, -FITC (clone 2E7; #121409, #121419), anti-mouse CD44-APC/Fire 750 (clone IM7; #103061), anti-mouse CD62L-Brilliant Violet 510 (clone MEL-14; 104441), anti-mouse CXCR6-Brilliant Violet 421 (clone SA051D1; #151109), anti-mouse CD11c Alex Fluor 647 (clone N418; #117314), anti-mouse CD11b-Alexa Fluor 488 (clone M1/70; #101219); Bioss: anti-mouse CXCL16 Alexa Fluor 488 (polyclonal; #bs-1441R-A488).

Techniques: Expressing, Luciferase, Isolation, Fluorescence, Staining

Journal: Stem Cell Research & Therapy

Article Title: LncRNA FTX/OCT4/miR-122-5p orchestrates self-renewal and lineage commitment of human dental pulp stem cells by directly targeting FOXO3

doi: 10.1186/s13287-025-04685-9

Figure Lengend Snippet: Characterization of hDPSCs. ( A – D ) The cellular morphologies of hDPSCs at passage 0 (P0), passage 3 (P3), passage 4 (P4), and passage 5 (P5) (×40) ( n = 3). ( E ) The expression levels of hDPSC surface markers (CD29, CD44, CD73, CD90, CD105, CD34 and CD45) were detected using flow cytometry. ( F ) hDPSCs possess highly proliferative capability confirmed by CCK-8 assay ( n = 3). ( G – I ) The osteogenic, adipogenic and chondrogenic differentiation capacity of hDPSCs were verified by Alizarin red (×100), oil red O (×200) and Alcian blue staining (×200), respectively ( n = 3). Error bars: mean ± SD

Article Snippet: The cells were first trypsinized and subsequently incubated in phosphate-buffered saline (PBS, Servicebio, China) with fluorescein-conjugated monoclonal antibodies specific for CD29-PE (Invitrogen, USA), CD44-APC-Cy7 (Tonbo Biosciences, USA), CD73-FITC (Invitrogen, USA), CD90-APC (Invitrogen, USA), CD105-PE (Invitrogen, USA), CD34-FITC (Invitrogen, USA) and CD45-PE-Cy7 (Invitrogen, USA) for 30 min, adhering strictly to the manufacturer’s instructions.

Techniques: Expressing, Flow Cytometry, CCK-8 Assay, Staining