Journal: Journal of Virology

Article Title: Adenovirus Modulates Toll-Like Receptor 4 Signaling by Reprogramming ORP1L-VAP Protein Contacts for Cholesterol Transport from Endosomes to the Endoplasmic Reticulum

doi: 10.1128/JVI.01904-16

Figure Lengend Snippet: Reagents and kits used in this study

Article Snippet: Description of antibodies used in the study a table ft1 table-wrap mode="anchored" t5 TABLE 2 caption a7 Reagent Company Catalog no. ALLN Calbiochem (Billerica, MA) 110044-82-1 Bio-Rad protein assay kit Bio-Rad (Hercules, CA) 500-0001 Cell fractionation kit Cell Biolabs AKR-171 HRS siRNA SMARTpool Dharmacon (Pittsburgh, PA) L-016835 ORP1L HRS siRNA SMARTpool Dharmacon M-008350 ORP5 siRNA SMARTpool Dharmacon L-009274 Rab7 siRNA SMARTpool Dharmacon L-010388 ECL detection reagent GE Life Sciences (Pittsburgh, PA) RPN2236 Percoll GE Life Sciences 17-0891-01 Amplex Red cholesterol assay kit Invitrogen (Grand Island, NY) {"type":"entrez-nucleotide","attrs":{"text":"A12216","term_id":"489486","term_text":"A12216"}} A12216 Lipofectamine Invitrogen 11668-019 Oligofectamine Invitrogen 12252-011 Bodipy 493/503 Invitrogen D3922 DAPI Invitrogen D1306 LipidTOX Invitrogen {"type":"entrez-nucleotide","attrs":{"text":"H34157","term_id":"979574","term_text":"H34157"}} H34157 SlowFade antifade kit Invitrogen S2828 Anti-GFP-agarose MBL (Woburn, MA) D153-8 Glutathione-agarose Pierce (Rockford, IL) 16100 AllStars negative-control siRNA Qiagen (Valencia, CA) 1027280 Cab-O-Sil Sigma (St. Louis, MO) 13760 Filipin Sigma F4767 Lipopolysaccharide (LPS) Sigma L5293 Poly- l -lysine Sigma P4707 Protein A Sepharose Sigma P3391 Open in a separate window Reagents and kits used in this study Cells and tissue culture.

Techniques: Cell Fractionation, Amplex Red Cholesterol Assay, Negative Control

Journal: PLoS Pathogens

Article Title: SopF, a phosphoinositide binding effector, promotes the stability of the nascent Salmonella -containing vacuole

doi: 10.1371/journal.ppat.1007959

Figure Lengend Snippet: (A) HeLa cells were transfected with a plasmid encoding for EGFP-SopF for 18 h and then subject to confocal fluorescence microscopy or sequential detergent fractionation. Left panel shows a representative confocal microscopy image. EGFP-SopF (greyscale), DNA (blue). Scale bar is 10 μm. Right panel shows immunoblotting analysis. Cells were collected and subject to sequential detergent fractionation. Equal volumes of saponin-soluble, TX-100-soluble, and SDS-soluble fractions were separated by SDS-PAGE and subject to immunoblotting with antibodies against GFP, Hsp27 (cytosol), calnexin (membranes) and lamin A/C (nucleus). Molecular mass markers are indicated on the left. Results are representative of two independent experiments. (B) As for (A) except HeLa cells were transfected with a plasmid encoding for FLAG-SopF. FLAG-SopF was detected by immunostaining (left panel) or immunoblotting (right panel) with anti-FLAG antibodies. (C) SopF partially colocalizes with actin-binding proteins found at cell adhesion sites. HeLa cells were transfected with pFLAG-SopF for 18 h, then fixed and immunostained with anti-FLAG, anti-moesin, anti-lamellipodin and anti-vasodilator-stimulated phosphoprotein (VASP) antibodies. Representative confocal microscopy images show FLAG-SopF in green and moesin, lamellipodin or VASP in red. Scale bars are 10 μm. Insets show enlargements of boxed areas.

Article Snippet: Membranes were blocked at room temperature for 1 h with Tris-buffered saline (TBS) containing 5% (w/v) skim milk powder and 0.1% (v/v) Tween-20 (TBST-milk), then incubated with the following primary antibodies overnight at 4˚C: mouse anti-FLAG M2 affinity isolated (1:2,000 dilution; Sigma), mouse anti-HA.11 ascites (1:2,000; BioLegend), rabbit polyclonal anti-GFP (1:40,000; Thermo), mouse anti-β-lactamase (clone 8A5.A10, 1:2,000 dilution; Thermo), mouse anti-Hsp27 (clone G31, 1:20,000; Cell Signaling), rabbit polyclonal anti-calnexin (1:40,000; Enzo), rabbit polyclonal anti-lamin A/C (1:5,000; Cell Signaling) or mouse anti-LAMP-1 (clone H4A3, 1:1,000 dilution; Developmental Studies Hybridoma Bank).

Techniques: Transfection, Plasmid Preparation, Fluorescence, Microscopy, Fractionation, Confocal Microscopy, Western Blot, SDS Page, Immunostaining, Binding Assay

Journal: PLoS Pathogens

Article Title: SopF, a phosphoinositide binding effector, promotes the stability of the nascent Salmonella -containing vacuole

doi: 10.1371/journal.ppat.1007959

Figure Lengend Snippet: (A) HeLa epithelial cells were infected with S . Typhimurium wild type (WT), Δ sopF , Δ sopF pSopF (comp), Δ sopF pSopF(1–345), Δ sopF pSopF(1–367) or Δ sopF pACYC177 (empty vector) bacteria. The proportion of cytosolic bacteria was determined by CHQ resistance assay at 90 min p.i. (upper panel) or GAL8 recruitment at 1 h p.i. (lower panel, all bacteria are constitutively expressing mCherry for fluorescence detection). Upper panel: data represent the mean ± SD (n≥3 independent experiments). Lower panel: Data represent the mean ± SD (total of >600 bacteria per strain from n≥3 independent experiments). Asterisks indicate data significantly different from WT infection (one-way ANOVA with Dunnett’s post-hoc test). (B) HeLa cells were transfected with plasmids encoding for FLAG-SopF(1–367) or FLAG-SopF(1–345) for 18 h. Cells were fixed and immunostained with anti-FLAG antibodies. DNA was stained with Hoechst 33342. Representative confocal microscopy images show FLAG-SopF in greyscale and DNA in blue. Scale bars are 10 μm. (C) Subcellular fractionation of transfected cells. HeLa cells were transfected with plasmids encoding for FLAG-SopF, FLAG-SopF(1–367) or FLAG-SopF(1–345) for 18 h, then collected and subjected to sequential detergent fractionation. Equal volumes of saponin-soluble, TX-100-soluble, and SDS-soluble fractions were separated by SDS-PAGE and subject to immunoblotting with antibodies against the FLAG epitope, Hsp27 (cytosol), calnexin (membranes) and lamin A/C (nucleus). Molecular mass markers are indicated on the left. Results are representative of two independent experiments. (D) C-terminal truncations of SopF lose plasma membrane association in the S . cerevisiae mss4 tet-off strain. Wild type (WT) and mss4 tet-off yeast strains were transformed with plasmids encoding for yEGFP-SopF, yEGFP-SopF C370S, yEGFP-SopF(1–367) or yEGFP-SopF(1–345) and the subcellular localization of SopF in live cells was visualized by widefield fluorescence microscopy. Representative fluorescence images are shown. Scale bars are 2 μm. The role of a potential lipidation site in SopF localization was assessed by site-directed mutagenesis of the Cys370 residue (C370S). The grey box depicts a domain of unknown function (DUF), DUF3626, spanning amino acid residues 178–338 of SopF. (E) Quantification of SopF localization in WT and mss4 tet-off yeast strains that were transformed and visualized as described in (D). Subcellular localization was categorized as cytosol, internal membrane sites (IMS), plasma membrane (PM), or IMS and PM. Results are expressed as the mean percentage of total yeast transformants (n = 300 cells from three independent transformations).

Article Snippet: Membranes were blocked at room temperature for 1 h with Tris-buffered saline (TBS) containing 5% (w/v) skim milk powder and 0.1% (v/v) Tween-20 (TBST-milk), then incubated with the following primary antibodies overnight at 4˚C: mouse anti-FLAG M2 affinity isolated (1:2,000 dilution; Sigma), mouse anti-HA.11 ascites (1:2,000; BioLegend), rabbit polyclonal anti-GFP (1:40,000; Thermo), mouse anti-β-lactamase (clone 8A5.A10, 1:2,000 dilution; Thermo), mouse anti-Hsp27 (clone G31, 1:20,000; Cell Signaling), rabbit polyclonal anti-calnexin (1:40,000; Enzo), rabbit polyclonal anti-lamin A/C (1:5,000; Cell Signaling) or mouse anti-LAMP-1 (clone H4A3, 1:1,000 dilution; Developmental Studies Hybridoma Bank).

Techniques: Infection, Plasmid Preparation, Expressing, Fluorescence, Transfection, Staining, Confocal Microscopy, Fractionation, SDS Page, Western Blot, FLAG-tag, Transformation Assay, Microscopy, Mutagenesis

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Control of Memory Phenotype T Lymphocyte Homeostasis: Role of Costimulation.

doi: 10.4049/jimmunol.2100653

Figure Lengend Snippet: FIGURE 1. Steady-state cycling of MP Tregs and CD41 MP T cells requires CD28 signaling in vivo. WT C57BL/6 mice were treated with Rat-IgG2a, anti-CD28 dAb, anti-CD80/86, CTLA4-Ig (abatacept), or anti-ICOS (250 mg/dose) on days 0, 2, 4, and 6 and spleens were harvested on day 8. (A) Represen- tative plot of Ly-6C versus Ki-67 expression on splenic Tregs. (B) Percentage of Ly6C−Tregs expressing Ki-67 after Ab treatments. (C) Absolute number of splenic CD41Foxp31 T cells after Ab treatments. (D) Representative plot of Ki-67 expression on CD41Foxp3−CD441 cells after Ab treatments. (E) Percent- age of CD41Foxp3−CD441 MP T cells expressing Ki-67. (F) Absolute number of CD41 MP T cells after Ab treatments. (G) Representative plot of Ki-67 expression on CD81 CM T cells. (H) Percentage of CD81CD62L1 T cells expressing Ki-67. (I) Absolute number of CD81 CM T cells after Ab treatments. (J) Representative plot of the percentage of CD81 EM cells expressing Ki-67. (K) Percentage of CD81 EM cells expressing Ki-67. (L) Absolute number of CD81 EM T cells after Ab treatments. (A)(L) represent the results of one experiment using two to five mice per group. ****p < 0.0001. ns, not significant.

Article Snippet: C57BL/6 mice were injected with following mAbs as indicated in the figure legends: 250 mg/dose mouse IgG1 (clone MOPC-21; Bio X Cell), mouse IgG2a (clone C1.18.4), rat IgG2a (clone 2A3; Bio X Cell), human CD28 domain-specific Ab (100 mg/dose; Bristol Myers Squibb), 250 mg/dose antiCD80 (clone 1G10; Bio X Cell), anti-CD86 (clone GL-1; Bio X Cell), CTLA4-Ig (abatacept, Bristol Myers Squibb), anti-ICOS (clone 7E.17G9; Bio X Cell), anti-CTLA4 (clone UC10-4F10-11; Bio X Cell), anti PD-1 (clone 29F.1A12; Bio X Cell), TIM-3 (clone RMT3.23; Bio X Cell), TIGIT (clone 1G9; Bio X Cell), BTLA (clone 6A6, Bio X Cell), VISTA (clone 13F3; Bio X Cell), CD70 (clone FR70; Bio X Cell), CD48 (clone HM48.1; Bio X Cell), TNF-a (clone XT3.11; Bio X Cell), IFN-g (clone XMG1.2; Bio X Cell), IFNAR (clone MAR1-5A3; Bio X Cell), IFN-g receptor (clone GR20), IL-7 (clone M25; Bio X Cell), IL-7Ra (clone A7R34; Bio X Cell), CD40 (50 mg/dose; clone FGK4.5; Bio X Cell, CD40L (clone MR-1; Bio X Cell), GITR (clone DTA-1; Bio X Cell), OX40 (clone OX-86; Bio X Cell), 4-1BB (clone LOB12.3; Bio X Cell), and anti MHC-I (500 mg/dose; clone M1/42; Bio X Cell).

Techniques: In Vivo, Expressing

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Control of Memory Phenotype T Lymphocyte Homeostasis: Role of Costimulation.

doi: 10.4049/jimmunol.2100653

Figure Lengend Snippet: FIGURE 2. CD80/CD86 signaling is critical for CD41 MP cell and Treg homeostasis in both lymphoid and nonlymphoid sites, but the effects of mAb treatment are reversible. (AH) WT C57BL/6 mice were injected with either rat IgG2a or anti-CD28 dAb every other day for 6 d, and lymphocytes from mesenteric lymph nodes (AD) and liver (E-H) were harvested on day 8. (A) Ki-67 expression on Ly-6C−. (B) Absolute number of Tregs on day 8. (C) Ki-67 expression on CD41Foxp3−CD441 T cells. (D) Absolute number of CD41Foxp3−CD441 T cells. (E) Ki-67 expression on Ly-6C−Tregs. (F) Absolute number of Tregs. (G) Ki-67 expression on CD41Foxp3−CD441 T cells. (H) Absolute number of CD41Foxp3−CD441 T cells. (IP) C57BL/6 mice were injected with rat IgG2a or anti-CD80/ CD86 on days 0, 2, 4, and 6. Splenic cells were harvested on day 30 or 45. (I and J) Percentage of CD41Foxp31 Tregs and Ki-67 expression on Ly-6C−Tregs on day 30. (K and L) Percentage of Tregs and Ki-67 expression on Ly-6C−Tregs on day 45. (M and N) Absolute number of Tregs and CD41Foxp3−CD441 T cells on day 30. (O and P) Absolute number of Treg and CD41Foxp3−CD441 T cells on day 45. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. ns, not significant.

Article Snippet: C57BL/6 mice were injected with following mAbs as indicated in the figure legends: 250 mg/dose mouse IgG1 (clone MOPC-21; Bio X Cell), mouse IgG2a (clone C1.18.4), rat IgG2a (clone 2A3; Bio X Cell), human CD28 domain-specific Ab (100 mg/dose; Bristol Myers Squibb), 250 mg/dose antiCD80 (clone 1G10; Bio X Cell), anti-CD86 (clone GL-1; Bio X Cell), CTLA4-Ig (abatacept, Bristol Myers Squibb), anti-ICOS (clone 7E.17G9; Bio X Cell), anti-CTLA4 (clone UC10-4F10-11; Bio X Cell), anti PD-1 (clone 29F.1A12; Bio X Cell), TIM-3 (clone RMT3.23; Bio X Cell), TIGIT (clone 1G9; Bio X Cell), BTLA (clone 6A6, Bio X Cell), VISTA (clone 13F3; Bio X Cell), CD70 (clone FR70; Bio X Cell), CD48 (clone HM48.1; Bio X Cell), TNF-a (clone XT3.11; Bio X Cell), IFN-g (clone XMG1.2; Bio X Cell), IFNAR (clone MAR1-5A3; Bio X Cell), IFN-g receptor (clone GR20), IL-7 (clone M25; Bio X Cell), IL-7Ra (clone A7R34; Bio X Cell), CD40 (50 mg/dose; clone FGK4.5; Bio X Cell, CD40L (clone MR-1; Bio X Cell), GITR (clone DTA-1; Bio X Cell), OX40 (clone OX-86; Bio X Cell), 4-1BB (clone LOB12.3; Bio X Cell), and anti MHC-I (500 mg/dose; clone M1/42; Bio X Cell).

Techniques: Injection, Expressing

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Control of Memory Phenotype T Lymphocyte Homeostasis: Role of Costimulation.

doi: 10.4049/jimmunol.2100653

Figure Lengend Snippet: FIGURE 7. M1/42 administration enhances systemic proinflammatory cytokine production. (AC) WT C57BL/6 mice were injected with rat IgG2a or M1/42 (pan-antiMHC-I) on days 0, 2, 4, and 6. Plasma was col- lected on day 8 to measure IFN-g (A), TNF-a (B), and IL-6 (C) levels by cytometric bead array.

Article Snippet: C57BL/6 mice were injected with following mAbs as indicated in the figure legends: 250 mg/dose mouse IgG1 (clone MOPC-21; Bio X Cell), mouse IgG2a (clone C1.18.4), rat IgG2a (clone 2A3; Bio X Cell), human CD28 domain-specific Ab (100 mg/dose; Bristol Myers Squibb), 250 mg/dose antiCD80 (clone 1G10; Bio X Cell), anti-CD86 (clone GL-1; Bio X Cell), CTLA4-Ig (abatacept, Bristol Myers Squibb), anti-ICOS (clone 7E.17G9; Bio X Cell), anti-CTLA4 (clone UC10-4F10-11; Bio X Cell), anti PD-1 (clone 29F.1A12; Bio X Cell), TIM-3 (clone RMT3.23; Bio X Cell), TIGIT (clone 1G9; Bio X Cell), BTLA (clone 6A6, Bio X Cell), VISTA (clone 13F3; Bio X Cell), CD70 (clone FR70; Bio X Cell), CD48 (clone HM48.1; Bio X Cell), TNF-a (clone XT3.11; Bio X Cell), IFN-g (clone XMG1.2; Bio X Cell), IFNAR (clone MAR1-5A3; Bio X Cell), IFN-g receptor (clone GR20), IL-7 (clone M25; Bio X Cell), IL-7Ra (clone A7R34; Bio X Cell), CD40 (50 mg/dose; clone FGK4.5; Bio X Cell, CD40L (clone MR-1; Bio X Cell), GITR (clone DTA-1; Bio X Cell), OX40 (clone OX-86; Bio X Cell), 4-1BB (clone LOB12.3; Bio X Cell), and anti MHC-I (500 mg/dose; clone M1/42; Bio X Cell).

Techniques: Injection, Clinical Proteomics

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Control of Memory Phenotype T Lymphocyte Homeostasis: Role of Costimulation.

doi: 10.4049/jimmunol.2100653

Figure Lengend Snippet: FIGURE 6. CD28 signaling modulates Treg and CD41 MP T cell proliferation after treatment with agonistic anti-CD40. WT C57BL/6 mice were treated with rat IgG2a, anti-CD28 dAb (100 mg/dose; i.p.), anti-CD40 (50 mg/dose; i.p.), anti-CD40L (250 mg/dose), or anti-CD40 together with anti-CD28 dAb on days 0, 2, 4, and 6 and spleens were harvested on day 8. (A) Representative plots of Ly-6C versus Ki-67 expression on CD41Foxp31 Tregs. (B) Ki-67 expression on Ly6C−Tregs after Ab treatment. (C) Absolute number of splenic CD41Foxp31 T cells after Ab treatment. (D) Expression of Ki-67 on CD41Foxp3−CD441 MP T cells after Ab treatment. (E) Percentage of Ki-67 expression on CD41 MP T cells. (F) Absolute number of CD41 MP T cells after Ab treatment. (G) Representative plot of the expression of Ki-67 on CD81 CM T cells. (H) Ki-67 expression on CD81 CM T cells. (I) Absolute number of CD81 CM T cells after Ab treatment. (J) Representative plot of the expression of Ki-67 on CD81 EM T cells. (K) Ki-67 expression on CD81 EM T cells. (L) Absolute number of CD81 EM T cells after Ab treatment. (A)(L) represent the result of one experiment of two using five mice per group. **p < 0.01, ***p < 0.001, ****p < 0.0001. ns, not significant.

Article Snippet: C57BL/6 mice were injected with following mAbs as indicated in the figure legends: 250 mg/dose mouse IgG1 (clone MOPC-21; Bio X Cell), mouse IgG2a (clone C1.18.4), rat IgG2a (clone 2A3; Bio X Cell), human CD28 domain-specific Ab (100 mg/dose; Bristol Myers Squibb), 250 mg/dose antiCD80 (clone 1G10; Bio X Cell), anti-CD86 (clone GL-1; Bio X Cell), CTLA4-Ig (abatacept, Bristol Myers Squibb), anti-ICOS (clone 7E.17G9; Bio X Cell), anti-CTLA4 (clone UC10-4F10-11; Bio X Cell), anti PD-1 (clone 29F.1A12; Bio X Cell), TIM-3 (clone RMT3.23; Bio X Cell), TIGIT (clone 1G9; Bio X Cell), BTLA (clone 6A6, Bio X Cell), VISTA (clone 13F3; Bio X Cell), CD70 (clone FR70; Bio X Cell), CD48 (clone HM48.1; Bio X Cell), TNF-a (clone XT3.11; Bio X Cell), IFN-g (clone XMG1.2; Bio X Cell), IFNAR (clone MAR1-5A3; Bio X Cell), IFN-g receptor (clone GR20), IL-7 (clone M25; Bio X Cell), IL-7Ra (clone A7R34; Bio X Cell), CD40 (50 mg/dose; clone FGK4.5; Bio X Cell, CD40L (clone MR-1; Bio X Cell), GITR (clone DTA-1; Bio X Cell), OX40 (clone OX-86; Bio X Cell), 4-1BB (clone LOB12.3; Bio X Cell), and anti MHC-I (500 mg/dose; clone M1/42; Bio X Cell).

Techniques: Expressing

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Control of Memory Phenotype T Lymphocyte Homeostasis: Role of Costimulation.

doi: 10.4049/jimmunol.2100653

Figure Lengend Snippet: FIGURE 8. CD28 signaling regulates NK celldependent enhancement of CD41 MP T cell, but not CD81 MP T cell, proliferation. C57BL/6 mice were treated with rat IgG2a, M1/42 (500 mg/dose), anti-CD80/86 (250 mg/dose), and M1/42 together with anti-CD80/86 on days 0, 2, 4, and 6 and spleens were harvested on day 8. (A) Representative plots of Ly-6C versus Ki-67 expression on CD41Foxp31 splenic Tregs. (B) Expression of Ki-67 on Ly6C−Tregs after Ab treatments. (C) Absolute number of splenic CD41Foxp31 T cells after Ab treatment. (D) Representative plots of the expression of Ki-67 on CD41Foxp3−CD441 MP T cells after Ab treatment. (E) Expression of Ki-67 on CD41 MP T cells. (F) Absolute number of CD41 MP T cells after Ab treatment. (G) Representative plots of the expression of Ki-67 on CD81 CM T cells. (H) Percentage of Ki-671CD81 CM T cells. (I) Absolute number of CD81 CM T cells after Ab treatment. (J) Representa- tive plots of Ki-67 expression on CD81 EM T cells. (K) Percentage of Ki-671CD81 CM T cells. (L) Absolute number of CD81 EM T cells after Ab treatment. (A)(L) represent the result of one experiment of two using five mice per group. ****p < 0.0001. ns, not significant.

Article Snippet: C57BL/6 mice were injected with following mAbs as indicated in the figure legends: 250 mg/dose mouse IgG1 (clone MOPC-21; Bio X Cell), mouse IgG2a (clone C1.18.4), rat IgG2a (clone 2A3; Bio X Cell), human CD28 domain-specific Ab (100 mg/dose; Bristol Myers Squibb), 250 mg/dose antiCD80 (clone 1G10; Bio X Cell), anti-CD86 (clone GL-1; Bio X Cell), CTLA4-Ig (abatacept, Bristol Myers Squibb), anti-ICOS (clone 7E.17G9; Bio X Cell), anti-CTLA4 (clone UC10-4F10-11; Bio X Cell), anti PD-1 (clone 29F.1A12; Bio X Cell), TIM-3 (clone RMT3.23; Bio X Cell), TIGIT (clone 1G9; Bio X Cell), BTLA (clone 6A6, Bio X Cell), VISTA (clone 13F3; Bio X Cell), CD70 (clone FR70; Bio X Cell), CD48 (clone HM48.1; Bio X Cell), TNF-a (clone XT3.11; Bio X Cell), IFN-g (clone XMG1.2; Bio X Cell), IFNAR (clone MAR1-5A3; Bio X Cell), IFN-g receptor (clone GR20), IL-7 (clone M25; Bio X Cell), IL-7Ra (clone A7R34; Bio X Cell), CD40 (50 mg/dose; clone FGK4.5; Bio X Cell, CD40L (clone MR-1; Bio X Cell), GITR (clone DTA-1; Bio X Cell), OX40 (clone OX-86; Bio X Cell), 4-1BB (clone LOB12.3; Bio X Cell), and anti MHC-I (500 mg/dose; clone M1/42; Bio X Cell).

Techniques: Expressing