human colon carcinoma cell line caco 2  (ATCC)

Journal: Foods

Article Title: Valorization and Functional Enhancement of Mature Assam Tea Leaves Through Indigenous Filamentous Fungi-Based Fermentation for Functional Drink Development

doi: 10.3390/foods15091562

Figure Lengend Snippet: The cytotoxicity of Caco-2 cells ( A ) and HT-29 cells ( B ) after treatment with co-fermented mature tea leaves samples for 72 h. Different lowercase letters indicate significant differences among fermentation times within the same treatment ( p < 0.05).

Article Snippet: Human colon carcinoma cell line (Caco-2) and Human colon adenocarcinoma cell line (HT-29) obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA) were used to carry out the cell cytotoxicity test.

Techniques:

Journal: iScience

Article Title: Cisplatin-incorporating gelatin-coated gadolinium oxide nanoparticles for cancer theranostics

doi: 10.1016/j.isci.2026.115010

Figure Lengend Snippet: In vitro cytotoxicity of gelatin-CDDP-Gd 2 O 3 NPs (A–E) Viability of HeLa (A), MCF-7 (B), A549 (C), HEK293 (D), and L929 (E) cells treated with gelatin-CDDP-Gd 2 O 3 NPs or CDDP alone at various CDDP concentrations (log scale). (F) The viability of HeLa cells treated with gelatin-CDDP-Gd 2 O 3 NPs in the presence of varying concentrations of TIMP-1 & 2 mixtures. Data are presented as mean ± SD ( n = 5). Statistical significance was evaluated using one-way ANOVA (∗ p < 0.05).

Article Snippet: L929 cells , ATCC , Cat# CCL-1.

Techniques: In Vitro

Journal: bioRxiv

Article Title: Systematic real-time profiling of Salmonella type III effector translocation provides quantitative resolution of the T3SS-1/T3SS-2 secretion dichotomy

doi: 10.64898/2026.03.06.710167

Figure Lengend Snippet: Representative HiBiT blots were obtained from three experimental conditions: (1) SPI-1–permissive LB conditions, including total lysates (TL) from early stationary phase cultures (OD 600 = 2.0) or overnight cultures, along with the corresponding secreted fractions (secretomes); (2) SPI-2–inducing conditions using mildly acidic LPM medium (pH 5.8); and (3) infection-derived fractions from HeLa cells infected at a multiplicity of infection (MOI) of 50, separated into TX-100–insoluble pellets (representing bacteria-associated effectors) and TX-100– soluble supernatants (representing translocated effectors). For each effector, a representative blot from a condition in which a clear HiBiT signal was obtained is shown. The accompanying six-square panel indicates detection (green), detection at higher protein load (orange), no detection (red), or not determined (black) across: (A) LB TL (OD 600 = 2.0 and/or LB O/N), (B) LB secretomes (O/N), (C) SPI-2–inducing conditions TL, (D) TX-100-insoluble pellet fraction (infected HeLa cells) TL, (E) TX-100-soluble supernatant (translocated effectors), and (F) real-time HiBiT translocation readout (infected HeLa cells). Representative blots obtained from LB OD 600 = 2.0 TL: AvrA (PVDL0187), GtgA (PVDL0313), GtgE (PVDL0246), SipB (PVDL0183), SipC (PVDL0028), SlrP (PVDL0184), SopB (PVDL0024), SopD (PVDL0185), SptP (PVDL0034), and SteC (PVDL0263). Representative blots obtained from LB O/N TL: CigR (PVDL0045), SifB (PVDL0041), SipA (PVDL0328), SopD2 (PVDL0299), SopE (PVDL0267), SopE2 (PVDL0033), SopF (PVDL0278), SpvB (PVDL0425), SseF (PVDL0257), SseK1 (PVDL0248), SseL (PVDL0249), SseM (PVDL0046), and SteB (PVDL0311). Representative blots obtained from SPI-2–inducing conditions: GogB (PVDL0298), PipB (PVDL0251), PipB2 (PVDL0040), SifA (PVDL0312), SpvC (PVDL0426), SpvD (PVDL0427), SrfJ (PVDL0280), SseD (PVDL0270), SseG (PVDL0258), SseI (PVDL0259), SseJ (PVDL0275), SseK2 (PVDL0260), SseK3 (PVDL0289), SspH2 (PVDL0261), and SteD (PVDL0271). Representative blots obtained from additional conditions: SopA from infection-derived Triton X-100–insoluble pellet fractions from infected HeLa cells (PVDL0032; 8 h post-infection (hpi)) and SteA (PVDL0029; LB O/N secretome).

Article Snippet: HeLa cells (human cervical adenocarcinoma, American Type Culture Collection, Manassas, VA, USA; ATCC ® CCL-2TM) and a stable HeLa cell line expressing LgBiT (HeLa-LgBiT) , kindly provided by Prof. Samuel Wagner, were maintained in GlutaMAX TM -supplemented Dulbecco’s Modified Eagle Medium (DMEM; Invitrogen, cat. no. 31966047) containing 10% foetal bovine serum (FBS; Invitrogen, cat. no. 10270106) and antibiotics (50 units/mL penicillin and 50 μg/mL streptomycin; Gibco, cat. no. 15070063).

Techniques: Infection, Derivative Assay, Bacteria, Translocation Assay

Journal: bioRxiv

Article Title: Systematic real-time profiling of Salmonella type III effector translocation provides quantitative resolution of the T3SS-1/T3SS-2 secretion dichotomy

doi: 10.64898/2026.03.06.710167

Figure Lengend Snippet: Representative HiBiT blots illustrating the secretion potential of all 39 Salmonella typedetected green fluorescent objects III effectors in overnight (O/N) LB cultures, which mainly supports SPI-1–dependent secretion. For each effector, total lysates (TL) are shown alongside their corresponding secretome (S) fractions, providing a comparative map of basal, SPI-1–mediated secretion capacity. TL samples are included to demonstrate expression relative to secretion under these conditions. As in , the accompanying six-square panel summarizes detection outcomes across the major experimental categories, with green indicating detection, orange detection at higher protein load, red no detection, and black not determined, across: (A) LB TL (OD 600 = 2.0 and/or LB O/N), (B) LB secretomes (O/N), (C) SPI-2–inducing conditions TL, (D) TX-100-insoluble pellet fraction (infected HeLa cells) TLs, (E) TX-100-soluble supernatants (translocated effectors), and (F) real-time HiBiT translocation readout (infected HeLa cells). All HiBiT-tagged effectors were expressed from endogenously modified strains in the GFP-expressing SL1344 background ( hisG 46 P tet :: gfp ; PVD0002), except for SipA, SpvB, SpvC, and SpvD, which were expressed in the wild-type SL1344 background (PVD0001). Individual strain identifiers are listed in Supplementary Table S2 .

Article Snippet: HeLa cells (human cervical adenocarcinoma, American Type Culture Collection, Manassas, VA, USA; ATCC ® CCL-2TM) and a stable HeLa cell line expressing LgBiT (HeLa-LgBiT) , kindly provided by Prof. Samuel Wagner, were maintained in GlutaMAX TM -supplemented Dulbecco’s Modified Eagle Medium (DMEM; Invitrogen, cat. no. 31966047) containing 10% foetal bovine serum (FBS; Invitrogen, cat. no. 10270106) and antibiotics (50 units/mL penicillin and 50 μg/mL streptomycin; Gibco, cat. no. 15070063).

Techniques: Expressing, Infection, Translocation Assay, Modification

Journal: bioRxiv

Article Title: Systematic real-time profiling of Salmonella type III effector translocation provides quantitative resolution of the T3SS-1/T3SS-2 secretion dichotomy

doi: 10.64898/2026.03.06.710167

Figure Lengend Snippet: HiBiT blotting analysis of 10 selected Salmonella type III effectors—SopA, SipC, SopE2, SteA, SopB, SptP, SseM, PipB2, SifB, and CigR—during infection of HeLa cells. CigR data are not shown due to the absence of a detectable HiBiT signal. (A) Representative HiBiT blots of Triton X-100–soluble (host cell cytosolic; translocated effectors) and Triton X-100–insoluble (bacteria-associated; expressed effectors) fractions collected at the indicated hours post-infection (hpi). GAPDH and GFP immunoblots are shown as representative and reproducible controls from the SopB experiment and were used as markers of the TX-100–soluble host cell fraction and as a normalization control for bacterial load, respectively. The black arrowhead denotes the mono-ubiquitinated form of the effector SopB, indicative of host-cell translocation. SopA, SipC, and SopE2 were detectable exclusively in the TX-100–insoluble fraction under the infection conditions tested (8–16 hpi), indicating bacterial expression but no detectable host cell translocation. (B) Densitometric quantification of HiBiT signals over time (background-corrected signal intensity; a.u.: arbitrary units). Upper panels depict translocated effector levels (tr.), quantified from the TX-100–soluble fraction. Middle panels show bacteria-associated effector levels (expr.), quantified from the TX-100–insoluble fraction. Lower graphs represent the relative translocation efficiency (tr./expr.), calculated as the ratio of translocated (TX-100–soluble) to bacteria-associated (TX-100–insoluble) HiBiT signal. Quantification is shown for those effectors displaying detectable host-cell translocation under the infection conditions tested (8–16 hpi) (SopB, SptP, SifB, PipB2, and SseM). HeLa cells were infected at an MOI of 50, and samples were processed as described in Methods. Abbreviations: tr., translocated effector (TX-100–soluble fraction); expr., bacteria-associated (expressed) effector (TX-100–insoluble fraction).

Article Snippet: HeLa cells (human cervical adenocarcinoma, American Type Culture Collection, Manassas, VA, USA; ATCC ® CCL-2TM) and a stable HeLa cell line expressing LgBiT (HeLa-LgBiT) , kindly provided by Prof. Samuel Wagner, were maintained in GlutaMAX TM -supplemented Dulbecco’s Modified Eagle Medium (DMEM; Invitrogen, cat. no. 31966047) containing 10% foetal bovine serum (FBS; Invitrogen, cat. no. 10270106) and antibiotics (50 units/mL penicillin and 50 μg/mL streptomycin; Gibco, cat. no. 15070063).

Techniques: Infection, Bacteria, Western Blot, Control, Translocation Assay, Expressing

Journal: bioRxiv

Article Title: Systematic real-time profiling of Salmonella type III effector translocation provides quantitative resolution of the T3SS-1/T3SS-2 secretion dichotomy

doi: 10.64898/2026.03.06.710167

Figure Lengend Snippet: HeLa cells stably expressing LgBiT were infected with a constitutively eGFP-expressing Salmonella enterica serovar Typhimurium SL1344 strain, either wild-type (WT eGFP+, left panels) or expressing chromosomally integrated sopB-HiBiT (SopB-HiBiT eGFP+, right panels), at multiplicities of infection (MOIs) of 10 (green), 50 (red), or 100 (blue). Infections were performed in quadruplicate, and real-time acquisition was conducted for 24 h using a multimode plate reader under environmental control. Data acquisition started 2 h post-infection (hpi). Top-to-bottom panels show: (i) fold change in HeLa cell count based on label-free automated brightfield imaging (10x objective); (ii) green fluorescence (RFU) as a proxy of bacterial load; (iii) average size (µm 2 ) of detected green fluorescent objects; and (iv) luminescence signal (RLU) either background levels or SopB-HiBiT translocation into LgBiT-expressing HeLa cells via NanoLuc complementation. Vivazine Live Cell Substrate was added following the gentamicin protection step and prior to acquisition to enable continuous luminescence detection throughout the infection course, starting at 2 hpi. Data points (every 30’) represent the mean ± SD of quadruplicates. WT-infected controls (left) confirm minimal background luminescence in the absence of HiBiT-tagged effectors.

Article Snippet: HeLa cells (human cervical adenocarcinoma, American Type Culture Collection, Manassas, VA, USA; ATCC ® CCL-2TM) and a stable HeLa cell line expressing LgBiT (HeLa-LgBiT) , kindly provided by Prof. Samuel Wagner, were maintained in GlutaMAX TM -supplemented Dulbecco’s Modified Eagle Medium (DMEM; Invitrogen, cat. no. 31966047) containing 10% foetal bovine serum (FBS; Invitrogen, cat. no. 10270106) and antibiotics (50 units/mL penicillin and 50 μg/mL streptomycin; Gibco, cat. no. 15070063).

Techniques: Stable Transfection, Expressing, Infection, Control, Cell Characterization, Imaging, Fluorescence, Translocation Assay

Journal: bioRxiv

Article Title: Systematic real-time profiling of Salmonella type III effector translocation provides quantitative resolution of the T3SS-1/T3SS-2 secretion dichotomy

doi: 10.64898/2026.03.06.710167

Figure Lengend Snippet: HeLa cells were infected with constitutively eGFP-expressing S. Typhimurium SL1344 wild-type (WT) or mutant strains deficient in key virulence factors: ΔinvA (T3SS-1-deficient), ΔssaV (T3SS-2-deficient), or a translational knockout of the SPI-2 effector SifA (ΔsifA, tko). Infections were performed at a multiplicity of infection (MOI) of 50, and real-time monitoring was conducted over 24 hours using automated label-free imaging in a multimodal plate reader under environmental control. Fluorescence parameters were normalized to their respective maxima (% max). Total green fluorescence (RFU, green) and average green object size (µm 2 , red) serve as proxies for intracellular bacterial load and bacterial distribution within infected cells, respectively. Data points represent mean ± SD of technical quadruplicates.

Article Snippet: HeLa cells (human cervical adenocarcinoma, American Type Culture Collection, Manassas, VA, USA; ATCC ® CCL-2TM) and a stable HeLa cell line expressing LgBiT (HeLa-LgBiT) , kindly provided by Prof. Samuel Wagner, were maintained in GlutaMAX TM -supplemented Dulbecco’s Modified Eagle Medium (DMEM; Invitrogen, cat. no. 31966047) containing 10% foetal bovine serum (FBS; Invitrogen, cat. no. 10270106) and antibiotics (50 units/mL penicillin and 50 μg/mL streptomycin; Gibco, cat. no. 15070063).

Techniques: Infection, Expressing, Mutagenesis, Knock-Out, Imaging, Control, Fluorescence

Journal: bioRxiv

Article Title: Systematic real-time profiling of Salmonella type III effector translocation provides quantitative resolution of the T3SS-1/T3SS-2 secretion dichotomy

doi: 10.64898/2026.03.06.710167

Figure Lengend Snippet: HeLa cells stably expressing LgBiT were infected with a panel of 39 endogenously HiBiT-tagged Salmonella enterica serovar Typhimurium SL1344 strains, all generated in a constitutively eGFP-expressing background ( hisG 46 P tet :: gfp ; PVD0002). Infections were performed at a multiplicity of infection (MOI) of 50, and real-time monitoring was conducted for 24 hours (from 2 to 26 hpi) using automated multimodal imaging in a plate reader under environmental control. Total green fluorescence (RFU, green) and average green object size (µm , red) serve as proxies for intracellular bacterial load and bacterial distribution within infected cells, respectively. Luminescence signal (RLU, blue) reflects NanoLuc complementation following translocation of HiBiT-tagged effectors into the host cell cytosol. Fluorescence and luminescence parameters were normalized to their respective maxima (% max) to facilitate comparison across strains. Data points represent mean ± SD of technical quadruplicates. Individual strain identifiers and effector annotations are listed in Supplementary Table S2 .

Article Snippet: HeLa cells (human cervical adenocarcinoma, American Type Culture Collection, Manassas, VA, USA; ATCC ® CCL-2TM) and a stable HeLa cell line expressing LgBiT (HeLa-LgBiT) , kindly provided by Prof. Samuel Wagner, were maintained in GlutaMAX TM -supplemented Dulbecco’s Modified Eagle Medium (DMEM; Invitrogen, cat. no. 31966047) containing 10% foetal bovine serum (FBS; Invitrogen, cat. no. 10270106) and antibiotics (50 units/mL penicillin and 50 μg/mL streptomycin; Gibco, cat. no. 15070063).

Techniques: Stable Transfection, Expressing, Infection, Generated, Imaging, Control, Fluorescence, Translocation Assay, Comparison

Journal: bioRxiv

Article Title: Systematic real-time profiling of Salmonella type III effector translocation provides quantitative resolution of the T3SS-1/T3SS-2 secretion dichotomy

doi: 10.64898/2026.03.06.710167

Figure Lengend Snippet: HeLa cells stably expressing LgBiT were infected with endogenously HiBiT-tagged Salmonella enterica serovar Typhimurium SL1344 effector strains in three genetic backgrounds: wild-type (WT) SL1344 carrying constitutive eGFP + ( hisG 46 P tet ::gfp; PVD0002; green), the T3SS-1–deficient ΔinvA::cat mutant (PVD0316; blue), and the T3SS-2–deficient ΔssaV::cat mutant (PVD0317; red). Luminescence profiles (RLU), reflecting NanoLuc complementation following effector translocation into the host cytosol, were recorded from 2 to 26 hours post-infection (hpi). Panels show a representative subset of effectors grouped by secretion system dependence—SPI-1 (SopB, SptP, SopD; left), SPI-1/2 (GogB [25% SPI-2 contribution], SseM [56%], SseJ [95%]; middle), and SPI-2 (SteD, SseF, SspH2; right)—as determined by phase-resolved AUC analysis (see Methods and Supplementary Table S5 ). The complete set of translocation profiles for all 39 effectors is shown in . Infections were performed at a multiplicity of infection (MOI) of 50, and real-time luminescence monitoring was conducted using a Spark Cyto 400 multimodal plate reader under environmental control in the presence of Nano-Glo Vivazine. Luminescence values were background-subtracted (non-infected HeLa). Data points represent mean ± SD of technical quadruplicates. Individual strain identifiers and effector annotations are provided in Supplementary Table S2.

Article Snippet: HeLa cells (human cervical adenocarcinoma, American Type Culture Collection, Manassas, VA, USA; ATCC ® CCL-2TM) and a stable HeLa cell line expressing LgBiT (HeLa-LgBiT) , kindly provided by Prof. Samuel Wagner, were maintained in GlutaMAX TM -supplemented Dulbecco’s Modified Eagle Medium (DMEM; Invitrogen, cat. no. 31966047) containing 10% foetal bovine serum (FBS; Invitrogen, cat. no. 10270106) and antibiotics (50 units/mL penicillin and 50 μg/mL streptomycin; Gibco, cat. no. 15070063).

Techniques: Stable Transfection, Expressing, Infection, Mutagenesis, Translocation Assay, Control

Journal: bioRxiv

Article Title: Systematic real-time profiling of Salmonella type III effector translocation provides quantitative resolution of the T3SS-1/T3SS-2 secretion dichotomy

doi: 10.64898/2026.03.06.710167

Figure Lengend Snippet: HeLa cells stably expressing LgBiT were infected with endogenously HiBiT-tagged Salmonella enterica serovar Typhimurium SL1344 effector strains in three genetic backgrounds: wild-type SL1344 carrying constitutive eGFP + ( hisG 46 P tet :: gfp ; PVD0002; green), the T3SS-1–deficient ΔinvA::cat mutant (PVD0316; blue), and the T3SS-2–deficient ΔssaV::cat mutant (PVD0317; red). Luminescence profiles (RLU), reflecting NanoLuc complementation following effector translocation into the host cytosol, were recorded from 2 to 26 hours post-infection (hpi). Infections were performed at a multiplicity of infection (MOI) of 50, and real-time luminescence monitoring was conducted using a Spark Cyto 400 multimodal plate reader under environmental control in the presence of Nano-Glo Vivazine. Luminescence values were background-subtracted (non-infected HeLa). Effectors are displayed alphabetically and distributed across panels (A–C). Data points represent mean ± SD of technical quadruplicates. Individual strain identifiers and effector annotations are provided in Supplementary Table S2 .

Article Snippet: HeLa cells (human cervical adenocarcinoma, American Type Culture Collection, Manassas, VA, USA; ATCC ® CCL-2TM) and a stable HeLa cell line expressing LgBiT (HeLa-LgBiT) , kindly provided by Prof. Samuel Wagner, were maintained in GlutaMAX TM -supplemented Dulbecco’s Modified Eagle Medium (DMEM; Invitrogen, cat. no. 31966047) containing 10% foetal bovine serum (FBS; Invitrogen, cat. no. 10270106) and antibiotics (50 units/mL penicillin and 50 μg/mL streptomycin; Gibco, cat. no. 15070063).

Techniques: Stable Transfection, Expressing, Infection, Mutagenesis, Translocation Assay, Control

Journal: bioRxiv

Article Title: Systematic real-time profiling of Salmonella type III effector translocation provides quantitative resolution of the T3SS-1/T3SS-2 secretion dichotomy

doi: 10.64898/2026.03.06.710167

Figure Lengend Snippet: HeLa cells stably expressing cytosolic LgBiT were infected with selected HiBiT-tagged S. Typhimurium SL1344 strains in a constitutively eGFP-expressing background ( his G46 P tet ::gfp; PVD0002), including T3SS-1–deficient ( ΔinvA; blue) and T3SS-2–deficient ( ΔssaV; red) derivatives. Luminescence was measured at 24 hours post-infection (hpi), corresponding to the indicated timepoint from continuous real-time acquisition using (A) furimazine, added shortly before readout, or (B) vivazine, present throughout infection. Bars represent raw luminescence values (RLU; mean ± SD of technical replicates). Strain identifiers and effector annotations are listed in Supplementary Table S2 .

Article Snippet: HeLa cells (human cervical adenocarcinoma, American Type Culture Collection, Manassas, VA, USA; ATCC ® CCL-2TM) and a stable HeLa cell line expressing LgBiT (HeLa-LgBiT) , kindly provided by Prof. Samuel Wagner, were maintained in GlutaMAX TM -supplemented Dulbecco’s Modified Eagle Medium (DMEM; Invitrogen, cat. no. 31966047) containing 10% foetal bovine serum (FBS; Invitrogen, cat. no. 10270106) and antibiotics (50 units/mL penicillin and 50 μg/mL streptomycin; Gibco, cat. no. 15070063).

Techniques: Stable Transfection, Expressing, Infection