Journal: Vaccine: X

Article Title: Secreted heat shock protein gp96-Ig and OX40L-Fc combination vaccine enhances SARS-CoV-2 Spike (S) protein-specific B and T cell immune responses

doi: 10.1016/j.jvacx.2022.100202

Figure Lengend Snippet: Characteristics of the cell line expressing gp96-Ig, SARS-CoV-2 Spike (S) protein and OX40L-Fc. Cell line (AD100) was transfected with plasmids encoding a) gp96-Ig and full length protein S and b) gp96-Ig, full length protein S and OX40L-Fc. c) Secreted gp96-Ig was measured in the cell supernatant by ELISA. One million cells were plated in 1 ml for 24 h. Purified gp96-Ig was used as standard d) Secreted OX40L-Fc was measured in the cell supernatant by ELISA. One million cells were plated in 1 ml for 24 h. Purified OX40L-Fc was used as standard e) SARS-CoV2 protein S expression was analyzed by immunofluorescence f) SARS-CoV2 protein S expression in supernatant was measured by ELISA. One million cells were plated in 1 ml for 48 h and purified SARS-CoV2 protein S was used as standard.

Article Snippet: One million cells were plated in 1 ml of growth medium for 24 h and secreted gp96-Ig production was determined by ELISA using sheep anti-human gp96 antibody (HSP90B1-R&D Systems, Cat. No. AF7606) as coating antibody, peroxidase conjugated goat anti-human IgG antibody as a secondary antibody (Abcam Ab7499), and purified gp96-Ig IgG1 as a standard.

Techniques: Expressing, Transfection, Enzyme-linked Immunosorbent Assay, Purification, Immunofluorescence

Journal: Vaccine: X

Article Title: Secreted heat shock protein gp96-Ig and OX40L-Fc combination vaccine enhances SARS-CoV-2 Spike (S) protein-specific B and T cell immune responses

doi: 10.1016/j.jvacx.2022.100202

Figure Lengend Snippet: Gp96-Ig and OX40L-Fc increase S protein specific IgG responses in vivo . a) C57Bl6 mice were vaccinated at day 0 and 14 with different concentrations of cell-based gp96-Ig vaccine that expressed SARS-CoV-2 glycoprotein S and OX40L-Fc or with AD100 or PBS (controls). b) Mice were vaccinated at day 0 and 14 with 1 μg/ml of ZVX-55 and ZVX-60 or with AD100 and PBS (controls). Serum was collected 5 days after last vaccination, and S protein specific IgG response was analyzed by ELISA. c) Mice were vaccinated at day 0 and 14 with 1 μg/ml ZVX-55 or ZVX-60 and S protein specific IgG response in serum was analyzed at day 5, 14 and 19. Data represent 3 independent biological replicates per group and mean ± standard error. To compare control (ZVX55) with experimental (ZVX60) group (alpha level of 0.05) unpaired t -test (two-tailed) was applied, *p < 0.05, **p < 0.01, and ***p < 0.001.

Article Snippet: One million cells were plated in 1 ml of growth medium for 24 h and secreted gp96-Ig production was determined by ELISA using sheep anti-human gp96 antibody (HSP90B1-R&D Systems, Cat. No. AF7606) as coating antibody, peroxidase conjugated goat anti-human IgG antibody as a secondary antibody (Abcam Ab7499), and purified gp96-Ig IgG1 as a standard.

Techniques: In Vivo, Enzyme-linked Immunosorbent Assay, Control, Two Tailed Test

Journal: Vaccine: X

Article Title: Secreted heat shock protein gp96-Ig and OX40L-Fc combination vaccine enhances SARS-CoV-2 Spike (S) protein-specific B and T cell immune responses

doi: 10.1016/j.jvacx.2022.100202

Figure Lengend Snippet: Gp96-Ig and OX40L-Fc induce B cells responses. C57Bl6 mice were vaccinated at day 0 and 14 with a cell-based gp96-Ig vaccine that expressed SARS-CoV-2 glycoprotein S (ZVX-55, 1ug gp96-Ig) and OX40L-Fc (ZVX-60,1 ug gp96-Ig) or with AD100 or PBS (controls). a) Spleen cells (SPL) were isolated from vaccinated and control mice 5 days after last vaccination, stained for CD45, CD3, CD19, IgM, CD21, CD23, CD49, CD93. Frequency of CD19 + IgM+ (activated B cells) and CD21 + CD23- (marginal zone, MZ), CD21 + CD23+ (follicular, FO) and CD21-CD23- (double negative or ABC cells) CD19 + IgM + cells was determined by flow cytometry. b) SPL were isolated from unvaccinated mice and co-cultured with vaccine cells (ZVX55 or ZVX60) and control cells AD100 at 5:1 ratio for 96 h. Frequency of activated B cells (CD19 + IgM + ) within total CD45 + T cells and frequency of FO (CD21 + CD23 + ) within CD19 + IgM + cells was determined by flow cytometry. Data represent 3 independent biological replicates per group and mean ± standard error. To compare > 2 experimental groups, 2-way analysis of variance (ANOVA) test with Holm-Sidak multiple-comparison test were applied, *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: One million cells were plated in 1 ml of growth medium for 24 h and secreted gp96-Ig production was determined by ELISA using sheep anti-human gp96 antibody (HSP90B1-R&D Systems, Cat. No. AF7606) as coating antibody, peroxidase conjugated goat anti-human IgG antibody as a secondary antibody (Abcam Ab7499), and purified gp96-Ig IgG1 as a standard.

Techniques: Isolation, Control, Staining, Flow Cytometry, Cell Culture, Comparison

Journal: Vaccine: X

Article Title: Secreted heat shock protein gp96-Ig and OX40L-Fc combination vaccine enhances SARS-CoV-2 Spike (S) protein-specific B and T cell immune responses

doi: 10.1016/j.jvacx.2022.100202

Figure Lengend Snippet: Gp96-Ig and OX40L-Fc induce T follicular helper (TFH) cell responses. C57Bl6 mice were vaccinated at day 0 and 14 with a cell-based gp96-Ig vaccine that expressed SARS-CoV-2 glycoprotein S (ZVX-55, 1ug gp96-Ig) and OX40L-Fc (ZVX-60,1 ug gp96-Ig) or with AD100 or PBS (controls). a) Spleen cells (SPL) were isolated from vaccinated and control mice 5 days after last vaccination, stained for CD45, CD3, CD4, PD1 and CXCR5. Frequency of PD1 + CXCR5+ (TFH cells) within CD4 + T cells was determined by flow cytometry. b) SPL were isolated from unvaccinated mice and co-cultured with vaccine cells (ZVX55 or ZVX60) and control cells AD100 at 5:1 ratio for 96 h. Frequency of TFH cells (PD1 + CXCR5 + ) within total CD4 + T cells was determined by flow cytometry. Data represent 3 independent biological replicates per group and mean ± standard error. To compare > 2 experimental groups, 2-way analysis of variance (ANOVA) test with Holm-Sidak multiple-comparison test were applied, *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: One million cells were plated in 1 ml of growth medium for 24 h and secreted gp96-Ig production was determined by ELISA using sheep anti-human gp96 antibody (HSP90B1-R&D Systems, Cat. No. AF7606) as coating antibody, peroxidase conjugated goat anti-human IgG antibody as a secondary antibody (Abcam Ab7499), and purified gp96-Ig IgG1 as a standard.

Techniques: Isolation, Control, Staining, Flow Cytometry, Cell Culture, Comparison

Journal: Vaccine: X

Article Title: Secreted heat shock protein gp96-Ig and OX40L-Fc combination vaccine enhances SARS-CoV-2 Spike (S) protein-specific B and T cell immune responses

doi: 10.1016/j.jvacx.2022.100202

Figure Lengend Snippet: Enhancement of S1- specific CD8 + T cell responses by Gp96-Ig-S-OX40L-Fc in the spleen, lung tissue, and BAL . a) 5 days after the vaccination of HLA-A2 transgenic mice (n = 3–5) with the ZVX-55, ZVX-60 vaccine cells (1ug secreted gp96-Ig) or AD100 or PBS (controls), splenocytes (SPL), lung cells and bronchioalveolar lavage (BAL) cells were isolated from vaccinated and control mice (PBS). Cells were stained with HLA-A2 02–01 pentamer containing YLQPRTFLL peptides, followed by surface staining for CD45, CD3, CD4, CD8, CD69, CXCR6. Bar graphs represent percentage of the pentamer positive cells within CD8 + T cells. b) 5 days after the vaccination of C57Bl6 mice (n = 3), splenocytes and lung cells were isolated from vaccinated and control mice (PBS and AD100) and in vitro restimulated with S1 and S2 overlapping peptides in the presence of protein transport inhibitor, brefeldin A for the last 5 h of culture. After 20 h of culture, intracellular cytokine (IFNg, TNFa and IL-2) staining was preformed to quantify protein S-specific CD8 + T-cell responses. Cytokine expression in the presence of no peptides was considered background and it was subtracted from the responses measured from peptide pool stimulated samples for each individual mouse. Data represent at least 2 technical replicates with 3–5 independent biologic replicates per group and mean ± standard error. To compare > 2 experimental groups, 2-way analysis of variance (ANOVA) test with Holm-Sidak multiple-comparison test were applied, *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: One million cells were plated in 1 ml of growth medium for 24 h and secreted gp96-Ig production was determined by ELISA using sheep anti-human gp96 antibody (HSP90B1-R&D Systems, Cat. No. AF7606) as coating antibody, peroxidase conjugated goat anti-human IgG antibody as a secondary antibody (Abcam Ab7499), and purified gp96-Ig IgG1 as a standard.

Techniques: Transgenic Assay, Isolation, Control, Staining, In Vitro, Expressing, Comparison

Journal: The Journal of Cell Biology

Article Title: Cerebellar ataxia disease–associated Snx14 promotes lipid droplet growth at ER–droplet contacts

doi: 10.1083/jcb.201808133

Figure Lengend Snippet: Snx14 localizes at ER–LD contacts after OA treatment. (A) Localization of Snx14 in U2OS cells in the absence and presence of OA, respectively. CoIF staining of the cells transiently expressing an untagged Snx14 were performed with α-Snx14 antibody (green), and α-HSP90B1 (ER marker, red) antibodies and LDs were stained with MDH (blue) and imaged by confocal microscopy. Inset of OA-treated cells displays Snx14 accumulating around LDs. Scale bar = 20 µm. Scale bar of insets = 5 µm. (B) Schematic diagram showing Snx14 EGFP-APEX2 fusion at ER–LD contacts. (C) TEM micrographs showing untreated and DAB + H 2 O 2 –treated cells following OA treatment. The dark precipitate with DAB + H 2 O 2 treatment indicates presence of Snx14 EGFP-APEX2 . Scale bar = 0.5 µm. (D) TEM micrograph showing SNX14 EGFP-APEX2 -expressing cell with DAB precipitate at ER–LD contacts. (D′) Image from D showing pseudocolored ER (red), LD (yellow), and green arrows pointing at DAB precipitate specifically at the junction of ER and LD. The blue arrows indicate a region of LD surface that lacks ER wrapping and also lacks DAB precipitate. Purple stars indicate LDs without any detectable ER association (and no DAB precipitate). The orange arrow indicates an ER membrane itself. Scale bar = 0.5 µm. (E) Lower-magnification TEM micrographs of a Snx14 EGFP-APEX2 -expressing cell following OA treatment and stained with DAB, showing precipitate surrounding various LDs entangled with the ER network. Scale bar = 0.5 µm.

Article Snippet: The primary antibodies used are mouse anti-Hsp90B1 (1:300; Sigma-Aldrich; AMAb91019), rabbit anti-Snx14 (HPA017639), rabbit anti-EGFP (1:300), rabbit anti-Flag (1:200; Sigma-Aldrich; F7425), mouse anti-ACSL3 (1:200; Novus Biologicals; H00002181-B01P).

Techniques: Staining, Expressing, Marker, Confocal Microscopy

Journal: The Journal of Cell Biology

Article Title: Cerebellar ataxia disease–associated Snx14 promotes lipid droplet growth at ER–droplet contacts

doi: 10.1083/jcb.201808133

Figure Lengend Snippet: Snx14 is topologically anchored in the ER and interacts with LDs in trans. (A) LD flotation assay of OA-treated U2OS cells expressing Snx14 EGFP . Lanes indicate postnuclear supernatant (PNS), cytosol, total membrane, and LD float fractions. (B) Schematic diagram of Snx14 fragment constructs tagged with EGFP. Snx14 FL depicts the full-length human Snx14. Snx14 N is the N-terminal fragment from the start that includes PXA and RGS domains. Snx14 PXCN includes the PX domain and C-Nexin domains. Snx14 PX consists of PX and Snx14 CN represents C-Nexin domain. An AH in the C-Nexin domain is identified as depicted in the schematic diagram. Snx14 PXCNΔH indicates the PX and C-Nexin domain from which the AH is deleted. Snx14 FLΔH depicts the full-length Snx14 with AH deletion. (C) Western blot showing distribution of Snx14 FL , Snx14 N , and Snx14 PXCN tagged with 3XFlag among total membrane and LD fractions following OA treatment. (D) U2OS cells transfected with Snx14 FL , Snx14 N , Snx14 PXCN , Snx14 PX , Snx14 CN , Snx14 PXCNΔH , and Snx14 FLΔH , respectively, and treated with OA for 16 h. CoIF staining with α−EGFP (green) and α-HSP90B1 (ER marker, red) and LDs stained with MDH (blue) and imaged by confocal microscopy. Scale bar = 20 µm. Inset scale bar = 5 µm. Cartoons represent the localization of the respective Snx14 fragments with respect to ER and LD.

Article Snippet: The primary antibodies used are mouse anti-Hsp90B1 (1:300; Sigma-Aldrich; AMAb91019), rabbit anti-Snx14 (HPA017639), rabbit anti-EGFP (1:300), rabbit anti-Flag (1:200; Sigma-Aldrich; F7425), mouse anti-ACSL3 (1:200; Novus Biologicals; H00002181-B01P).

Techniques: Expressing, Construct, Western Blot, Transfection, Staining, Marker, Confocal Microscopy

Journal: The Journal of Cell Biology

Article Title: Cerebellar ataxia disease–associated Snx14 promotes lipid droplet growth at ER–droplet contacts

doi: 10.1083/jcb.201808133

Figure Lengend Snippet: Loss of Snx14 perturbs LD size and morphology but does not change neutral lipid levels. (A) Confocal micrographs of WT and SNX14 KO cells treated with OA overnight. LDs visualized by MDH (black) and nucleus stained with Syto 85 orange fluorescent stain (blue outline). Images were processed so that LDs were converted to grayscale and inverted. Scale bar = 25 µm. (B) Quantification of average area covered by LDs per cell of representative images from A. Total LD area was derived from more than five fields of view, each consisting of approximately five cells or more of two different sets of experiments (total no. of cells >75; ***, P < 0.0001 unpaired t test with α = 0.05). (C) TEM micrographs of WT and SNX14 KO cells treated with OA overnight to visualize LD distribution and morphology. The top panels are lower magnification (scale bar = 2 µm). The bottom panels are higher magnification (scale bar = 1 µm). (D) Scatter dot plot of cross-sectional areas of LDs in WT and SNX14 KO cells as in C. Total LDs = 896; ***, P < 0.0001, Kolmogorov–Smirnov D test with α = 0.05. (E) Rescue of LD morphology in SNX14 KO cells by readdition of empty vector (EV), Snx14 FL , Snx14 N , Snx14 PXCN , Snx14 PXCNΔH , and Snx14 FLΔH , respectively, all tagged with EGFP. Cells were coIF stained with α-EGFP (green), α-HSP90B1 (ER, red), and LDs stained with MDH (blue) and imaged with confocal microscope. Scale bar = 50 µm. (F) Area covered by LDs in each cell from E analyzed and plotted. Total no. of cells quantified are 45 from two different sets of experiments (***, P < 0.0001, one-way ANOVA with α = 0.05). Line bars indicate mean ± SD.

Article Snippet: The primary antibodies used are mouse anti-Hsp90B1 (1:300; Sigma-Aldrich; AMAb91019), rabbit anti-Snx14 (HPA017639), rabbit anti-EGFP (1:300), rabbit anti-Flag (1:200; Sigma-Aldrich; F7425), mouse anti-ACSL3 (1:200; Novus Biologicals; H00002181-B01P).

Techniques: Staining, Derivative Assay, Plasmid Preparation, Microscopy

Journal: The Journal of Cell Biology

Article Title: Cerebellar ataxia disease–associated Snx14 promotes lipid droplet growth at ER–droplet contacts

doi: 10.1083/jcb.201808133

Figure Lengend Snippet: Snx14 functions independently of ER–LD protein, Seipin. (A) Confocal micrographs showing localization of Snx14 in WT and SEIPIN KO SUM159 cells. The cells were transfected with Snx14 EGFP and treated with OA overnight. Labels were Snx14 EGFP (α-EGFP), ER (α-HSP90B1), and LDs (MDH). Scale bar = 20 µm. Inset scale bar = 5 µm. (B) Test of rescue of LD morphology in SEIPIN KO examined by ectopic expression of Snx14 EGFP into SEIPIN KO . Labels were Snx14 EGFP (α-EGFP), ER (α-HSP90B1), and LDs (MDH stained, converted to grayscale, and inverted by ImageJ). Scale bar = 20 µm. (C) Area covered by LDs in each cell from B analyzed and plotted. Total no. of cells quantified is 23 from two different sets of experiments (***, P < 0.0001, one-way ANOVA with α = 0.05). Line bars indicate mean ± SD. (D) Test of rescue of LD morphology in SNX14 KO examined by readdition of Seipin mCherry and comparing to that of WT and SNX14 KO . Blue line depicts cell boundary. Black speckles represent LDs that are MDH-stained, converted to grayscale, and inverted by ImageJ. Scale bar = 20 µm. (E) Area covered by LDs in each cell from D analyzed and plotted. Total no. of cells quantified is 45 from two different sets of experiments (***, P < 0.0001, one-way ANOVA with α = 0.05). Line bars indicate mean ± SD. (F) Confocal micrographs of U2OS cells to examine localization of Seipin mCherry in WT cells, those transfected with Snx14 EGFP , and in SNX14 KO cells. Cells were transfected with Seipin-mCherry and treated with OA overnight. Labels were Snx14 EGFP (α-EGFP), Seipin-mCherry (α-mCherry), and LDs (MDH). Scale bar = 20 µm.

Article Snippet: The primary antibodies used are mouse anti-Hsp90B1 (1:300; Sigma-Aldrich; AMAb91019), rabbit anti-Snx14 (HPA017639), rabbit anti-EGFP (1:300), rabbit anti-Flag (1:200; Sigma-Aldrich; F7425), mouse anti-ACSL3 (1:200; Novus Biologicals; H00002181-B01P).

Techniques: Transfection, Expressing, Staining

Journal: The Journal of Cell Biology

Article Title: Cerebellar ataxia disease–associated Snx14 promotes lipid droplet growth at ER–droplet contacts

doi: 10.1083/jcb.201808133

Figure Lengend Snippet: Snx14 localizes at ACSL3-positive preLDs following OA addition. (A) Confocal micrographs of immunofluorescently stained Snx14 EGFP -expressing cells following OA treatment for t = 0, 1, 2, 4, 8, and 16 h. Labels were Snx14 EGFP (α-EGFP), ER (α-HSP90B1), and LDs (MDH). Scale bar = 2 µm. Line scans show the spatial distribution of Snx14 (green) with respect to ER (red) and LDs (blue). (B) Confocal micrographs of immunofluorescently stained Snx14 EGFP -expressing cells following OA treatment for t = 0, 1, 2, 4, 8, and 16 h. Labels were Snx14 EGFP (α-EGFP), native ACSL3 (α-ACSL3), and LDs (MDH). Scale bar = 2 µm. Line scans show the spatial distribution of Snx14 (green) with respect to ACSL3 (red) and LDs (blue). Scale bar = 2 µm. (C) Quantification of Pearson’s coefficient between Snx14 and ER of n = ∼20 cells depicted in A. (D) Quantification of Pearson’s coefficient between ACSL3 and LDs, and Snx14 and LDs of n = ∼20 cells depicted in B. (E) Quantification of Pearson’s coefficient between Snx14 and ACSL3 of n = ∼20 cells depicted in B. (F) Confocal micrographs examining localization of ACSL3 in WT and SNX14 KO cells treated with OA overnight. Labels were native ACSL3 (α-ACSL3, green) and LDs (MDH, magenta). Scale bar = 20 µm. (G) Confocal micrographs to examine localization of Snx14 in WT cells treated with negative scrambled siRNA (neg Si) or ACSL3-targeted siRNA (ACSL3 Si), respectively. The cells were transfected with Snx14 EGFP along with the respective siRNAs and treated with OA overnight. Labels were Snx14 EGFP (α-EGFP), ER (α-HSP90B1), and LDs (MDH). Scale bar = 20 µm. Inset scale bar = 5 µm.

Article Snippet: The primary antibodies used are mouse anti-Hsp90B1 (1:300; Sigma-Aldrich; AMAb91019), rabbit anti-Snx14 (HPA017639), rabbit anti-EGFP (1:300), rabbit anti-Flag (1:200; Sigma-Aldrich; F7425), mouse anti-ACSL3 (1:200; Novus Biologicals; H00002181-B01P).

Techniques: Staining, Expressing, Transfection

Journal: Journal of Healthcare Engineering

Article Title: GP96 and SMP30 Protein Priming of Dendritic Cell Vaccination Induces a More Potent CTL Response against Hepatoma

doi: 10.1155/2022/2518847

Figure Lengend Snippet: Lentiviral vectors: LV5: GP96 lentiviral vector structure; LV8: SMP30 lentiviral vector structure. (a) GP96-transfected DCs. (b) SMP30-transfected DCs. (a) and (b) show the same location.

Article Snippet: We used mouse anti-human HSP90, rabbit anti-human GP96 (Wuhan Boster Biological Technology Ltd.), and mouse anti-human SMP30 (Santa Cruz Biotechnology Inc.) antibodies.

Techniques: Plasmid Preparation, Transfection

Journal: Journal of Healthcare Engineering

Article Title: GP96 and SMP30 Protein Priming of Dendritic Cell Vaccination Induces a More Potent CTL Response against Hepatoma

doi: 10.1155/2022/2518847

Figure Lengend Snippet: WB after transfection. (a) Detection of GP96 in 6 groups. (b) Detection of SMP30 in 6 groups. (c) GAPDH as the internal control. The GP96 group expressed more GP96, and the SMP30 group expressed more SMP30, indicating that transfection was successful.

Article Snippet: We used mouse anti-human HSP90, rabbit anti-human GP96 (Wuhan Boster Biological Technology Ltd.), and mouse anti-human SMP30 (Santa Cruz Biotechnology Inc.) antibodies.

Techniques: Transfection, Control

Journal: Journal of Healthcare Engineering

Article Title: GP96 and SMP30 Protein Priming of Dendritic Cell Vaccination Induces a More Potent CTL Response against Hepatoma

doi: 10.1155/2022/2518847

Figure Lengend Snippet: Liver cancer model. (a) The liver cancer model. (b) The GP96 + SMP30 group: tumors had disappeared in some mice on day 4. (c) Tumors harvested from mice. (d) Negative TUNEL staining for apoptosis in the DC group. (e) Positive TUNEL staining for apoptosis in the GP96 + SMP30 group.

Article Snippet: We used mouse anti-human HSP90, rabbit anti-human GP96 (Wuhan Boster Biological Technology Ltd.), and mouse anti-human SMP30 (Santa Cruz Biotechnology Inc.) antibodies.

Techniques: TUNEL Assay, Staining

Journal: Journal of Healthcare Engineering

Article Title: GP96 and SMP30 Protein Priming of Dendritic Cell Vaccination Induces a More Potent CTL Response against Hepatoma

doi: 10.1155/2022/2518847

Figure Lengend Snippet: Tumor growth curve and ELISA results. Curve: the tumor volume in the GP96 + SMP30 group was smaller than that in the protein group on day 4, day 14, and day 16, P < 0.05. IL-2: the GP96 + SMP30 group showed secretion of more IL-2 than the other groups, P < 0.01. IFN- γ : the GP96 + SMP30 group showed secretion of more IFN- γ than the other groups, P < 0.01.

Article Snippet: We used mouse anti-human HSP90, rabbit anti-human GP96 (Wuhan Boster Biological Technology Ltd.), and mouse anti-human SMP30 (Santa Cruz Biotechnology Inc.) antibodies.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Journal of Healthcare Engineering

Article Title: GP96 and SMP30 Protein Priming of Dendritic Cell Vaccination Induces a More Potent CTL Response against Hepatoma

doi: 10.1155/2022/2518847

Figure Lengend Snippet: Immunohistochemical staining of tumors. (a) Tumor expressing IL-2 in the GP96 + SMP30 group; the brown cytoplasm indicates positive staining. (b) Tumor expressing IL-2 in the GP96 group; the colorless cytoplasm indicates negative staining. (c) Tumor expressing IFN- γ in the GP96 + SMP30 group; the brown cytoplasm indicates positive staining. (d) Tumor expressing IFN- γ in the SMP30 group; the colorless cytoplasm indicates negative staining.

Article Snippet: We used mouse anti-human HSP90, rabbit anti-human GP96 (Wuhan Boster Biological Technology Ltd.), and mouse anti-human SMP30 (Santa Cruz Biotechnology Inc.) antibodies.

Techniques: Immunohistochemical staining, Staining, Expressing, Negative Staining