human sclc cell lines h446  (ATCC)

Journal: Biomedicines

Article Title: TAF1 Suppresses MHC-I Expression and Correlates with Poor Immunotherapy Response in Small Cell Lung Cancer

doi: 10.3390/biomedicines14050973

Figure Lengend Snippet: TAF1 represses MHC-I expression in SCLC. ( A ) Correlation between TAF1 expression and MHC-I gene expression in the IMpower133 RNA-seq dataset; ( B ) Correlation between TAF1 expression and HLA-C protein levels in the TU-SCLC proteomic dataset; ( C ) Western blot analysis of TAF1 expression in DMS53 cells with or without TAF1 knockdown; β-actin was used as a loading control; ( D ) Flow cytometric analysis of HLA-A/B/C expression in DMS53-sh TAF1 and DMS53-shCtrl cells; ( E ) qPCR analysis of representative HLA expression ( HLA-A , HLA-B , HLA-C ) in DMS53-sh TAF1 versus DMS53-shCtrl cells; ( F ) Western blot analysis of TAF1 expression in NCI-H146 cells with or without TAF1 knockdown; β-actin was used as a loading control; ( G ) Flow cytometric analysis of HLA-A/B/C expression in NCI-H146-sh TAF1 and NCI-H146-shCtrl cells; ( H ) Western blot analysis of TAF1 expression in RP cells with or without Taf1 knockdown; β-actin was used as a loading control; ( I ) Flow cytometric analysis of H2Kb/Db expression in RP-sh Taf1 and RP-shCtrl cells; ( J ) Top five downregulated pathways in tumors with high TAF1 expression compared to those with low TAF1 expression in the IMpower133 cohort; ( K ) Top five downregulated pathways enriched in tumor cells with high TAF1 expression compared with low TAF1 expression in Rudin et al. cohort.

Article Snippet: Human SCLC cell lines NCI-H146 (HTB-173) and DMS53 (CRL-2062) were obtained from American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Expressing, Gene Expression, RNA Sequencing, Western Blot, Knockdown, Control

Journal: iScience

Article Title: UHRF1 drives subtype-independent aggressiveness and immune evasion in small cell lung cancer through PRC2 interactions

doi: 10.1016/j.isci.2026.115475

Figure Lengend Snippet: UHRF1 is broadly overexpressed in SCLC and inversely correlates with RB1 protein expression (A) UHRF1 mRNA and protein expression, derived from datasets GSE60052 and HRA003419, were compared between human SCLC tumors and normal lung tissues. For GSE60052 normal lung n = 7, SCLC n = 79. For HRA003419 normal lung n = 106, SCLC n = 107. ∗∗ p < 0.01 and ∗∗∗∗ p < 0.0001 by unpaired t test with Welch’s correction. (B) Correlation analysis of RB1 and UHRF1 protein expression demonstrating a significant inverse relationship, consistent with UHRF1 acting downstream of RB1 loss. (C) Kaplan-Meier overall survival analysis of patients with SCLC stratified by high (red) vs. low (blue) UHRF1 mRNA expression. Survival curves were analyzed using log rank (Mantel-Cox) test. p = 0.0205. (D) UHRF1 mRNA expression across human lung cancer cell lines, including 60 SCLC lines (red), from the DepMap Public 25Q3 database. ∗∗∗∗ p < 0.0001 by Spearman correlation. (E) Overview of RB1 and TP53 mutation status (top), and expression profiles of NE markers (SYP, CHGA, NCAM1, and INSM1) and non-NE markers (MYC and REST), lineage-specific transcription factors (ASCL1, NEUROD1, POU2F3, and YAP1), and UHRF1 across three SCLC cell lines used in this study (H526, H446, and DMS79), three non-NE SCLC cell lines (DMS114, SW1271, and H1048), and one non-malignant lung epithelial cell line (SALE). (F) Western blot analysis of UHRF1 protein levels in normal lung fibroblast (MRC5) and three NE SCLC cell lines. Actin was used as a loading control. Semi-quantitative values of UHRF1 protein intensity relative to MRC5 are shown in numbers below.

Article Snippet: Human SCLC cells (NCI-H526, NCI-H446 and DMS79) and MRC5 fibroblasts were obtained from ATCC.

Techniques: Expressing, Derivative Assay, Mutagenesis, Western Blot, Control

Journal: iScience

Article Title: UHRF1 drives subtype-independent aggressiveness and immune evasion in small cell lung cancer through PRC2 interactions

doi: 10.1016/j.isci.2026.115475

Figure Lengend Snippet: UHRF1 promotes proliferation, anchorage-independent growth, migration, and invasion in SCLC cells (A) Western blot analysis of UHRF1 expression in vector control (VC) and UHRF1 knockout (KO) cells across three SCLC cell lines. (B) Growth proliferation curves measured by AlamarBlue assay comparing NCI-H526 VC (black line; doubling time = 1.42 days) and UHRF1 KO (gray line; doubling time = 2.68 days). (C) Representative images of EdU incorporation assay (proliferating cells; red) in NCI-H526 VC and UHRF1 KO cells. Nuclei were counterstained with DAPI (blue). Scale bars, 50 μm. (D) Quantification of the proportion of EdU-positive cells across all three SCLC cell lines. ( n = 3). (E) Representative soft agar colony formation images from NCI-H526 VC and UHRF1 KO. (F) Quantification of colony numbers for each of the SCLC cell lines in the soft agar assy. (G) Representative images from scratch wound healing assays at 0 and 24 h in NCI-H446 VC, UHRF1 KO, and UHRF1-rescued cells. Scale bars, 500 μm. (I) Quantification of wound closure in the migration assay ( n = 15). (J) Representative images from Transwell invasion assays with crystal violet-stained cells (purple) in NCI-H446 VC, UHRF1 KO, and UHRF1-rescued groups. (K) Quantification of cells invaded in the transwell assay ( n = 6). For all histograms, each bar represents the mean ± SD; ns = not significant, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001 by unpaired two-tailed t test.

Article Snippet: Human SCLC cells (NCI-H526, NCI-H446 and DMS79) and MRC5 fibroblasts were obtained from ATCC.

Techniques: Migration, Western Blot, Expressing, Plasmid Preparation, Control, Knock-Out, Alamar Blue Assay, Staining, Transwell Assay, Two Tailed Test

Journal: iScience

Article Title: UHRF1 drives subtype-independent aggressiveness and immune evasion in small cell lung cancer through PRC2 interactions

doi: 10.1016/j.isci.2026.115475

Figure Lengend Snippet: UHRF1 promotes tumor growth and metastatic spread in SCLC xenograft models All results represent the NCI-H526 cell line. (A) Scheme of the orthotopic xenograft mouse model. (B) Representative images of lungs and primary lung tumors harvested at the study endpoint. (C) Comparison of primary tumor weights between vector control (VC; black) and UHRF1 KO (red) groups. Bars represent mean ± SD ( n = 7). ∗∗ p < 0.01 by unpaired two-tailed t test. (D) Representative bioluminescent imaging of mice and dissected major organs from the orthotopic model. (E) Heatmap-style binary presence (gray) or absence (white) chart summarizes the frequency and distribution of metastasis across orthotopic xenografts. (F) Quantification of the number of metastatic sites observed across orthotopic xenografts. VC in black; UHRF1 KO in red. Bars represent mean ± SD ( n = 7). ∗∗ p < 0.01 by unpaired two-tailed t test. (G) Schematic of the NCI-H526 intracardiac injection xenograft model. (H) Representative bioluminescent images of mice and major organs from the intracardiac model. (I) Heatmap illustrates the distribution of metastases across organs in the intracardiac xenograft model. (J) Quantification of the number of metastatic organ sites in each group. VC in black; UHRF1 KO in red. Bars represent mean ± SD ( n = 3). ∗∗ p < 0.01 by unpaired two-tailed t test. (K) Kaplan-Meier survival analysis show overall survival probabilities for the orthotopic xenograft model for VC (black) and UHRF1 KO (red). Survival curves were analyzed using the log rank (Mantel–Cox) test. p = 0.0381.

Article Snippet: Human SCLC cells (NCI-H526, NCI-H446 and DMS79) and MRC5 fibroblasts were obtained from ATCC.

Techniques: Comparison, Plasmid Preparation, Control, Two Tailed Test, Imaging, Injection

Journal: iScience

Article Title: UHRF1 drives subtype-independent aggressiveness and immune evasion in small cell lung cancer through PRC2 interactions

doi: 10.1016/j.isci.2026.115475

Figure Lengend Snippet: UHRF1 expression is associated with the NE phenotype in SCLC (A and B) Pearson’s correlation (r) analyses between UHRF1 expression and canonical markers of (A) NE and (B) non-NE phenotypes using human SCLC tumor data ( GSE60052 ). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001 by two-tailed Pearson’s correlation. (C and D) Gene Set Enrichment Analysis (GSEA) compares (C) NE signature enrichment and (D) non-NE signature enrichment between NCI-H526 vector control (VC) and UHRF1 KO orthotopic tumors. (E) NE score comparison between NCI-H526 VC and UHRF1 KO tumors. ∗∗∗∗ p < 0.0001 by two-tailed unpaired t test. (F) Representative immunohistochemistry images of NE markers (SYP, NCAM1, CHGA) and non-NE markers (REST, YAP1) in VC and UHRF1 KO tumors. Scale bars, 50 μm. (G) Quantifications of immunohistochemistry signal intensities. Bars represent mean +/- SD; VC (n = 7), UHRF1 KO (n = 6). ns = not significant, ∗ p < 0.05, ∗∗ p < 0.01 by unpaired two-tailed t test. (H) Western blot analysis shows the expression of UHRF1, NE markers (synaptophysin, NCAM1), non-NE markers (REST, YAP1), and actin as a loading control in NCI-H526 VC and UHRF1 KO tumor samples. Relative average protein abundance shown below each blot. (I and J) Pearson’s correlation analyses between EZH2 and UHRF1 expression in (I) clinical SCLC dataset HRA003419 and (J) GEMM tumors. (K and L) Western blot analysis of EZH2 levels in UHRF1 KO versus VC tumors derived from (K) NCI-H526 and (L) DMS79 xenografts. Actin was used as a loading control. Relative average protein abundance is shown below each blot.

Article Snippet: Human SCLC cells (NCI-H526, NCI-H446 and DMS79) and MRC5 fibroblasts were obtained from ATCC.

Techniques: Expressing, Two Tailed Test, Plasmid Preparation, Control, Comparison, Immunohistochemistry, Western Blot, Quantitative Proteomics, Derivative Assay

Journal: iScience

Article Title: UHRF1 drives subtype-independent aggressiveness and immune evasion in small cell lung cancer through PRC2 interactions

doi: 10.1016/j.isci.2026.115475

Figure Lengend Snippet: Loss of UHRF1 leads to more immune cell infiltration (A) Gene Set Enrichment Analysis (GSEA) comparing inflammatory response signature enrichment between NCI-H526 vector control (VC) and UHRF1 KO orthotopic tumors. (B) Representative IHC images for F4/80 macrophage staining. Scale bars, 50 μm; and, (C) signal quantification on Tp53/Rb1 (TR) DKO and Tp53/Rb1/Uhrf1 (TRU) TKO GEMM tumors. Bars represent mean ± SD ( n = 5). ∗ p < 0.05 by unpaired two-tailed t test. (D) Representative IHC images for CD3 T cell staining. Scale bars, 50 μm; and (E) signal quantification on GEMM tumors. Bars represent mean ± SD ( n = 5). ∗ p < 0.05 by unpaired two-tailed t test. (F) Western blot shows UHRF1 protein levels in primary mouse SCLC cells generated from GEMM tumors. (G–N) Quantification of various immune cell populations in TR DKO ( n = 14) and TRU TKO ( n = 9) tumor allografts. Bars represent mean ± SD. ns = not significant, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001 by unpaired two-tailed t test. (O) Quantification of the chemokines immunoassay analysis ( n = 2 independent batches). Bars represent mean ± SD ∗ p < 0.05 by unpaired two-tailed t test. (P) Western blot analysis of MAGE-A4 in VC and UHRF1 KO cells of all three SCLC cell lines. (Q) Methylation-specific PCR (MSP) analysis of specific CpG loci on MAGE-A4 in VC and UHRF1 KO cells of all three SCLC cell lines. M: methylated. U: unmethylated. (R) Quantification of MAGE-A4 MSP. (S) RT-qPCR analysis of MAGE-A4 expression in 5-azacytidine (5aza) treated and untreated (DMSO) VC and UHRF1 KO cells of all three SCLC cell lines. Bars represent mean ± SD.

Article Snippet: Human SCLC cells (NCI-H526, NCI-H446 and DMS79) and MRC5 fibroblasts were obtained from ATCC.

Techniques: Plasmid Preparation, Control, Staining, Two Tailed Test, Western Blot, Generated, Methylation, Quantitative RT-PCR, Expressing

Journal: Nature Communications

Article Title: Differential KEAP1/NRF2 mediated signaling widens the therapeutic window of redox-targeting drugs in SCLC therapy

doi: 10.1038/s41467-026-71608-4

Figure Lengend Snippet: A Left panel: The expression of 15 ACB genes for each SCLC ( n = 50) and NSCLC cell lines ( n = 98). Right panel: EC 50 values of selected SCLC ( n = 13) and NSCLC ( n = 32) cell lines. Dots represent mean EC 50 values derived from two (H1105, HCC33, HCC15, H1573, H1792, H1437, H3122, and H1651) or at least three independent experiments for the remaining cell lines, each performed with biological triplicates. Statistical significance was assessed using a two-tailed unpaired t- test. The exact p values are indicated in the figure. Cell lines with known loss-of-function mutations in KEAP1 or Cul3 are indicated as black dots. B The heatmap with the protein levels of SLC7A11, GSR, and TXN, analyzed in total cell extracts by immunoblotting. Cell lines with loss-of-function mutations (LOF) in KEAP1 or Cul3 are indicated with asterix. Protein expression in H1944 was set to 1 for each protein and for each experiment. Relative data represent mean of two (SLC7A11, GSR, TXN1 in SCLC; SLC7A11 in NSCLC) or three (GSR and TXN1 in NSCLC) independent experiments. C Expression values of ACB genes were normalized to normal lung tissue. Box plots display the median (center line), interquartile range (box), and whiskers extending to 1.5× the interquartile range; individual data points are overlaid. Statistical differences among groups were assessed using the Kruskal–Wallis test followed by two-sided Dunn’s multiple comparisons test comparing each tumor type with SCLC. Exact p values are indicated in the figure when significant (<0.05). Sample sizes for each group are indicated on the x -axis labels. D , E Data are presented as mean of EC 50 values of three independent experiments, each performed in biological replicate. E Statistical significance was assessed using a two-tailed unpaired t -test. Exac t p values are indicated in the figure when significant (<0.05). D CN: derived from chemo-naïve patients, PC: derived from patients after cisplatin plus etoposide therapy. E H69 CPR , H526 CPR , H209 CPR are cisplatin-resistant sublines. Source data are provided as a file.

Article Snippet: Human SCLC suspension cell lines (ATCC: NCI-H69 HTB-119, NCI-H82 HTB-175, NCI-H526 CRL-5811, NCI-H209 HTB-172, NCI-H1105 CRL-5856, NCI-H187 CRL-5804, DMS79 CRL-2049, NCI-H146 HTB-173, NCI-H2171 CRL-5929, NCI-H1963 CRL-5982, NCI-H378 CRL-5808, Beas-2B CRL-3588; DSMZ: HCC33 487), SCLC-16HC and the NSCLC cell line (ATCC: NCI-H1944 CRL-5907) were cultivated in RPMI-1640 (Gibco).

Techniques: Expressing, Derivative Assay, Two Tailed Test, Western Blot

Journal: Nature Communications

Article Title: Differential KEAP1/NRF2 mediated signaling widens the therapeutic window of redox-targeting drugs in SCLC therapy

doi: 10.1038/s41467-026-71608-4

Figure Lengend Snippet: A The NRF2 protein level in total cell extracts was analyzed in SCLC and non-cancerous cell lines upon treatment with DMSO (control) or CDDO-Me by immunoblotting. NRF2 expression in DMSO-treated H82 cells was set to 1 for each experiment. Relative data represent mean ± SD of two independent experiments for H2171, H146, DMS79, H378, 16HC, H187, and HCC33, and at least three independent experiments for the other cell lines. Statistical significance was assessed using a two-tailed unpaired t -test. Exact p values are indicated in the figure when significant (<0.05). B The induction of NQO1 and AKR1C3 transcripts upon CDDO-Me treatment in different cell lines was determined by expression profiling ( n = 3, mean ± SD, two-tailed unpaired t- test). Exact p values are indicated in the figure when significant (<0.05). C The induction of NQO1 and AKR1C3 proteins upon CDDO-Me (50 nM) treatment was determined by immunoblotting (representative of three independent experiments). D The top 45 genes highly correlated with resistance to TXNRD1 inhibition were analyzed by expression profiling of two non-cancerous and two SCLC cell lines treated with CDDO-Me compared to a DMSO control ( n = 3). ACBs are labeled in blue. E The most upregulated pathways upon CDDO-Me treatment according to a pathway enrichment analysis based on mRNA expression profiling data using Ingenuity Pathway Analysis (IPA, QIAGEN Inc., Redwood City, CA, USA). Canonical pathway enrichment was assessed using right-tailed Fisher’s exact test, and p value were adjusted for multiple testing where applicable. F Oxidized and reduced levels of PRDX1 protein were analyzed by immunoblotting in cells treated first with CDDO-Me (50 nM) for 24 h and then with the indicated concentrations of DKFZ-682 for 3 h. Bar diagrams summarize the quantitative results from independent experiments ( n = 2 for H526, H82 and Beas-2B, n = 4 for HaCaT, mean ± SD). Statistical significance was assessed using a two-tailed unpaired t -test. Exact p values are indicated in the figure when significant (<0.05). G The cells were treated with CDDO-Me (50 nM) or DMSO (control). After 24 h, a concentration series of DKFZ-682 was added for another 24 h and the cell viability was measured by the CellTiter-Glo assay. The fold change of EC 50 for DKFZ-682 upon CDDO-Me compared to cells without CDDO-Me pretreatment was calculated. Source data are provided as a file.

Article Snippet: Human SCLC suspension cell lines (ATCC: NCI-H69 HTB-119, NCI-H82 HTB-175, NCI-H526 CRL-5811, NCI-H209 HTB-172, NCI-H1105 CRL-5856, NCI-H187 CRL-5804, DMS79 CRL-2049, NCI-H146 HTB-173, NCI-H2171 CRL-5929, NCI-H1963 CRL-5982, NCI-H378 CRL-5808, Beas-2B CRL-3588; DSMZ: HCC33 487), SCLC-16HC and the NSCLC cell line (ATCC: NCI-H1944 CRL-5907) were cultivated in RPMI-1640 (Gibco).

Techniques: Control, Western Blot, Expressing, Two Tailed Test, Inhibition, Labeling, Concentration Assay, Glo Assay

Journal: Nature Communications

Article Title: Differential KEAP1/NRF2 mediated signaling widens the therapeutic window of redox-targeting drugs in SCLC therapy

doi: 10.1038/s41467-026-71608-4

Figure Lengend Snippet: Human SCLC cell line H209 was transplanted subcutaneously into nude mice (for details see method section Animal experiments). Tumor-bearing mice were distributed into groups ( n = 10) and treated with vehicle (black), cisplatin/etoposide (gray, chemo, receiving 3 weekly cycles of cisplatin 3 mg/kg, interperitonally, i.p., on Monday and etoposide 7.5 mg/kg, i.p., on Wednesday and Friday) or daily injections of DKFZ-608 (green, starting with dose escalation of 5 - > 15 mg/kg in the first 5 days, continued with 15 mg/kg up to day 40). One group (blue) started on the chemo regimen for 3 weeks followed by DKFZ-608 for 40 days. The tumor size ( A ), survival ( B ) and body weight ( C ) were monitored during the course of the experiment. In ( A ), the mean values ± SD of the tumor size are shown and the group receiving DKFZ-608 mono-therapy (green) is presented as two separate lines in order to allow a better presentation of animals reaching full remission. The comparison of survival groups ( B ) was calculated with log-rank (Mantel-Cox) test. Tumor growth ( A ) and body weight ( C ), presented as mean ± SD, were analyzed using linear mixed-effects models with restricted maximum likelihood estimation (REML) via the lme4 package in R. The model included treatment (categorical), time (continuous; days), and their interaction as fixed effects, with random intercepts for individual animals to account for repeated measurements. Post-hoc pairwise comparisons between treatments were conducted using estimated marginal means in the emmeans package, with Tukey-adjusted p values. Source data are provided as a file.

Article Snippet: Human SCLC suspension cell lines (ATCC: NCI-H69 HTB-119, NCI-H82 HTB-175, NCI-H526 CRL-5811, NCI-H209 HTB-172, NCI-H1105 CRL-5856, NCI-H187 CRL-5804, DMS79 CRL-2049, NCI-H146 HTB-173, NCI-H2171 CRL-5929, NCI-H1963 CRL-5982, NCI-H378 CRL-5808, Beas-2B CRL-3588; DSMZ: HCC33 487), SCLC-16HC and the NSCLC cell line (ATCC: NCI-H1944 CRL-5907) were cultivated in RPMI-1640 (Gibco).

Techniques: Comparison

Journal: Nature Communications

Article Title: Differential KEAP1/NRF2 mediated signaling widens the therapeutic window of redox-targeting drugs in SCLC therapy

doi: 10.1038/s41467-026-71608-4

Figure Lengend Snippet: A As a tumor model, NSG mice were engrafted subcutaneously with the human SCLC cell line H526 (for details see “Method” section Animal experiments). After CDDO-Me pre-treatment, the NSG mice were daily injected intraperitoneally with DKFZ-682, gradually increasing the dose from 4 mg/kg to 10 mg/kg that was then maintained for up to 3 weeks. The tumor size was monitored daily for reaching the humane end-points. B Mice were randomly distributed into groups ( n = 10) that were daily injected with vehicle, CDDO-Me (3 mg/kg), a low dose of DKFZ-682 (dose escalation from 1 mg/kg to 4 mg/kg), CDDO-Me (3 mg/kg) and a low dose of DKFZ-682 (4 mg/kg), CDDO-Me (3 mg/kg) and a high dose of DKFZ-682 (dose escalation from 4 mg/kg to 10 mg/kg). The plot shows the survival of individual groups (compated by the log-rank (Mantel-Cox) test). C , D Impact of CDDO-Me and DKFZ-682 treatment on TXNRD1 activity and protein levels in tumor and liver tissue derived from treated mice (mean ± SD; n = 6 animals per group, measured in technical duplicates). The residual enzymatic activity of TXNRD1 in resected tumor and liver tissue was assessed by the activity probe TRFS-Green. Significance of differences was calculated with a two-tailed unpaired t -test. E The pathways upregulated by CDDO-Me were analyzed in mouse tissues and tumors, performing RNAseq. Samples from tumor-bearing mice (H526 tumors) that were injected with CDDO-Me (3 mg/kg) for 4 days were compared to vehicle ( n = 5). Pathway enrichment analysis was performed using Ingenuity Pathway Analysis (IPA, QIAGEN Inc., Redwood City, CA, USA). Canonical pathway enrichment was assessed using right-tailed Fisher’s exact test, and p values were adjusted for multiple testing where applicable. F The tumor size measured 12 days after the daily mouse treatment was started ( n = 10, one-way ANOVA p = 0.0025; pairwise differences compared by two-tailed unpaired t -test). G The sensitivity to DKFZ-682 was tested in explanted tumors after mouse treatment for the indicated time. Tumor cells were treated with a concentration series of DKFZ-682 for 24 h and cell viability was quantified using CellTiter-Glo. Each dot represents a tumor from one mouse measured in a triplicate. Source data are provided as a file.

Article Snippet: Human SCLC suspension cell lines (ATCC: NCI-H69 HTB-119, NCI-H82 HTB-175, NCI-H526 CRL-5811, NCI-H209 HTB-172, NCI-H1105 CRL-5856, NCI-H187 CRL-5804, DMS79 CRL-2049, NCI-H146 HTB-173, NCI-H2171 CRL-5929, NCI-H1963 CRL-5982, NCI-H378 CRL-5808, Beas-2B CRL-3588; DSMZ: HCC33 487), SCLC-16HC and the NSCLC cell line (ATCC: NCI-H1944 CRL-5907) were cultivated in RPMI-1640 (Gibco).

Techniques: Injection, Activity Assay, Derivative Assay, Two Tailed Test, RNA sequencing, Concentration Assay